Treatment of metabolic disorders in canine animals

ABSTRACT

The present invention relates to one or more SGLT2 inhibitors or pharmaceutically acceptable forms thereof for use in the treatment and/or prevention of a metabolic disorder in a canine animal, preferably wherein the metabolic disorder is one or more selected from the group consisting of: ketoacidosis, pre-diabetes, insulin dependent diabetes mellitus, insulin resistance diabetes, insulin resistance, obesity, hyperglycemia, hyperglycemia induced cataract formation, impaired glucose tolerance, hyperinsulinemia, dyslipidemia, dysadipokinemia, subclinical inflammation, systemic inflammation, low grade systemic inflammation, hepatic lipidosis, inflammation of the pancreas, metabolic disorder consequences, such as hypertension, renal dysfunction and/or muscoskeletal disorders, and/or Syndrome X (metabolic syndrome), wherein preferably the development of hyperglycemia induced cataract formation is prevented or remission is achieved and/or wherein preferably the development of metabolic disorder consequences, such as hypertension, renal dysfunction and/or muscoskeletal disorders, is prevented or progression is slowed or remission is achieved.

FIELD OF THE INVENTION

The present invention relates to veterinary medicine, in particular tothe treatment and/or prevention of metabolic disorders in canineanimals.

BACKGROUND OF THE INVENTION

Canine animals, e.g. dogs, are affected by various metabolic disorders.A number of metabolic disorders are known in canine animals, includinghyperglycaemia, insulin resistance, diabetes, hepatic lipidosis,obesity, hyperinsulinaemia, impaired glucose tolerance, ketosis (inparticular ketoacidosis), dyslipidaemia, dysadipokinemia, subclinicalinflammation or systemic inflammation, in particular low grade systemicinflammation, which also comprises adipose tissue, Syndrome X (metabolicsyndrome) and/or inflammation of the pancreas. Various correlationsexist amongst these disorders. Among these disorders, in the dog,diabetes, in particular pre-diabetes and insulin dependent diabetesmellitus, as well as hyperglycaemia, insulin resistance and obesity aregaining more and more importance. This can at least partially beascribed to changing living and feeding behaviour and that companionanimals are living longer due to improved preventive veterinary careduring the last years.

Diabetes mellitus is characterized by disturbances in carbohydrate,protein and triglyceride metabolism based on a relative or absolute lackof insulin.

It is a relatively common endocrinopathy in canine animals like the dog.The incidence for diabetes in dogs has increased in the last decades toapproximately up to 1.0%. Several risk factors have been identified:age, obesity, neutering, gender and breed.

The current classification divides diabetes mellitus in humans intothree classes:

(1.) Type 1 which results from the loss of function of insulin secretingcells, e.g. by immunologic destruction of beta cells or insulinauto-antibodies (juvenile diabetes in humans);

(2.) Type 2 which results from a failure of the insulin stimulated cellsto respond properly to insulin stimuli; it is also associated to e.g.amyloid accumulation in beta cells; type 2 usually develops during along time of the so called pre-diabetes state;(3.) secondary diabetes mellitus which can due to diabetogenic drugs(e.g. long-acting glucosteroids, megestrol acetat, etc.) or to otherprimary diseases like pancreatitis, pancreas adenocarcinoma, cushing,hypo- or hyperthyroidism, growth-hormone producing tumors resulting inacromegaly.

Canine diabetes is not easily classified, although there are clearsimilarities and differences between the human and canine diseases.There is no evidence of a canine equivalent to type 2 diabetes, despiteobesity being as much a problem in pet dogs as it is in their owners.

The disease can be broadly divided into insulin deficiency diabetes andinsulin resistance diabetes (Catchpole et al., Diabetologia 2005. 48:1948-1956). Insulin deficiency is the most common type. In contrast tothe human situation it is not commonly found in young dogs, but ratherhas possibly similarities to the latent/late autoimmune diabetes of theadult (LADA) form of type 1 diabetes in man, which is characterised byprogressive beta cell destruction by autoimmune reactions.

Autoimmunity in dogs is however controversial. As antibodies have beendetected only in a subset of dogs with canine diabetes and are discussedto be a consequence rather than a cause of the disease (Catchpole etal., Diabetologia 2005. 48: 1948-1956).

Additionally, in intact female dogs a dioestrus/gestational dependentinsulin resistance diabetes is frequent.

For the treatment of diabetes in humans, especially of type 2 diabetesmellitus, several oral antihyperglycaemic drugs are approved. Thesedrugs act, e.g. by stimulating pancreatic insulin secretion in aglucose-independent or glucose-dependent manner(sulfonylurea/meglitinidcs, or DPP IV inhibitors, respectively), byenhancing tissue sensitivity to insulin (biguanides,thiazolidinediones), or by slowing postprandial intestinal glucoseabsorption (alpha-glucosidase inhibitors).

Some oral antihyperglycaemic drugs have been employed, but are eithernot effective in diabetic dogs e.g. sulfonylurea drugs or did show someeffects on glycemic control, but are unfavorable due to high prevalenceof adverse effects e.g. alpha-glucosidase inhibitors (Nelson et al. Jsmall Anim Pract 2000, 41, 486-490).

Other approaches have been contemplated for treating diabetes and reducehyperglycemia, including inhibition of the renal sodium-dependentglucose co-transporter SGLT2. SGLT2 in the kidney regulates glucoselevels by mediating the reabsorption of glucose back into the plasmafollowing filtration of the blood. SGLT2 inhibition thus inducesglycosuria and may reduce blood glucose levels. For example, compound1-cyano-2-(4-cyclopropyl-benzyl)-4-(β-D-glucopyranos-1-yl)-benzene isdescribed as an SGLT2 inhibitor in WO 2007/128749. A large variety offurther SGLT2 inhibitors are also known. In WO 2011/117295, which isconcerned with the medication of predominantly carnivorous non-humananimals with dipeptidyl peptidase IV (DPP-IV) inhibitors, various SGLT2inhibitors are recited amongst numerous other types of compounds in thecontext of combination therapies with DPP-IV inhibitors.

SGLT2 inhibition has not previously been contemplated for treatment ofmetabolic disorders in canine animals, such as dogs. In canine animals,medications for metabolic disorders are far less advanced than inhumans. Unfortunately, even if a treatment or prophylaxis is effectivein humans, e.g., or other non-canine animals, it is not possible toconclude that the same approach will also be effective, safe andotherwise appropriate in a canine animal, such as a dog.

Canine animals differ significantly from humans or, e.g., othercarnivores as cats in respect of their metabolisms.

Consequently, the pathophysiology of canine metabolic disorders, andthus also their responses to medication of such disorders differs fromother species.

Dogs display obesity and all characteristics of a metabolic syndromesimilar to e.g. humans and also cats. In contrast to these species, incanine animals this syndrome does not progress to a type 2 diabetes. Apathophysiological hallmark of type 2 diabetes in humans as well as infelines—the pancreatic islet amyloid deposition is absent in dogs(Verkest, Vet J, in press doi.org/10.1016/j.tvjl.2013.09.057)

Diabetic complication e.g. vision problems are commonly seen withdiabetes mellitus in dogs, but are rarely found in feline animals.Though, retinopathy is frequently detected in human diabetics—in dogs itis rarely found, but vision problems arise from keratopathy andespecially cataracts. These are encountered in up to 80% of diabeticdogs (Beam et al. Vet. Ophtalmol. 1999. 2, 169-172)

Optimal glycaemic control has been shown to be crucial to prevent thedevelopment or progression of cataracts (Wang et al. J Diabet. Compl. inpress, doi:0.1016/j.jdiacomp.2013.11.002)

The gold-standard treatment of diabetic dogs is currently considered tobe injection of insulin. However, no single type of insulin is routinelyeffective in maintaining control of glycaemia, even with twice-dailyadministration. Even regulated diabetics may eventually reach a pointwhere their blood glucose is no longer controlled and the insulin mustbe adjusted, whether by dose or type.

Also with strict compliance from the owner control is often poor andsecondary problems are common. Many owners find it impossible to achieveacceptable levels of compliance, as synchronization of food intake andinsulin injection is impossible in the majority of cases. Ultimatelymany dogs with diabetes mellitus are euthanized because of the disease.

The factors governing patient and owner compliance are also verydifferent. In dogs, oral administration, e.g., is yet more highlydesirable than in humans.

A treatment that would allow better compliance and therefore betterglycaemic control than current insulin-based treatments would help toattenuate the progression of the disease and delay or prevent onset ofcomplications in many animals.

No satisfactory treatment is currently available for metabolic disorderssuch as obesity, insulin resistance, hyperglycaemia, hyperinsulinaemia,impaired glucose tolerance, hepatic lipidosis, dyslipidaentia,dysadipokinemia, subclinical inflammation or systemic inflammation, inparticular low grade systemic inflammation, which also comprises adiposetissue, and associated disorders, such as Syndrome X (metabolicsyndrome). Furthermore, these metabolic disorders can be associated toor induced by hypo- or hyperthyroidism, hypercortisolism(hyperadrenocorticism, coshing) and/or growth-hormone access(acromegaly). These metabolic disorders might become clinically manifeste.g. by hypertension, cardiomyopathy, renal dysfunction and/ormusculoskeletal disorders in canine animals.

Thus, there remains a particular need for effective, safe and otherwiseappropriate treatments of metabolic disorders, including diabetes, incanine animals.

DISCLOSURE OF THE INVENTION Summary of the Invention

The present inventors have surprisingly found that inhibition of SGLT2is effective and safe in the treatment and/or prevention of metabolicdisorders in canine animals.

The present invention thus provides the use of one or more SGLT2inhibitors or a pharmaceutically acceptable form thereof in thetreatment and/or prevention of a metabolic disorder of a canine animal.

Further, the present invention provides the use of one or more SGLT2inhibitors or a pharmaceutically acceptable form thereof in thetreatment and/or prevention of a metabolic disorder of a canine animal,wherein the one or more SGLT2 inhibitors is1-cyano-2-(4-cyclopropyl-benzyl)-4-(β-D-glucopyranos-1-yl)-benzene(which is referred to in the following as compound A) or apharmaceutically acceptable form thereof.

Compound A has the following chemical formula:

Further aspects of the invention are defined below as well as in theclaims.

The pharmaceutically acceptable form of the one or more SGLT2inhibitors, preferably compound A, may be a crystalline complex betweenthe one or more SGLT2 inhibitors and one or more amino acids, such asproline.

According to the invention, the one or more SGLT2 inhibitors, preferablycompound A, or pharmaceutically acceptable form thereof may be provided,e.g., for oral or parenteral administration, preferably for oraladministration.

The one or more SGLT2 inhibitors, preferably compound A, or apharmaceutically acceptable form thereof may be administered in dosagesof 0.1 to 3.0 mg/kg body weight per day, preferably from 0.2 to 2.0mg/body weight per day, more preferably from 0.1 to 1 mg/body weight perday. Thus, the one or more SGLT2 inhibitors, preferably compound A, orpharmaceutically acceptable form thereof may be prepared for theadministration of 0.1 to 3.0 mg/kg body weight per day, preferably from0.2 to 2.0 mg/kg body weight per day, more preferably from 0.1 to 1mg/kg body weight per day.

The one or more SGLT2 inhibitors, preferably compound A, orpharmaceutically acceptable form thereof is preferably administered onlyonce per day.

The present invention also provides a pharmaceutical compositioncomprising one or more SGLT2 inhibitors, preferably compound A, or apharmaceutically acceptable form thereof, for use according to theinvention as disclosed herein.

In the examples provided herein, therapeutic and/or prophylacticbenefits resulting from inhibition of SGLT2 according to the presentinvention are demonstrated experimentally. Experimental data disclosedherein are intended to illustrate the invention, but not to have anylimiting effect upon the scope of protection, which is defined hereinbelow by the claims.

In particular, the present inventors have surprisingly found that theuse of one or more SGLT2 inhibitors, preferably compound A, according tothe present invention advantageously leads to a reduction ofhyperglycaemia and/or additionally to an improved e.g. 9 or 24 hglycaemic profile in a hyperglycaemic (e.g. diabetic) canine. Thus, thiscan result in a reduction of insulin needed to treat diabetic canineanimals.

As the absorption and onset of action (glycosuria) is very fast andprominent (Examples 1 and 2) treating a canine with a newly diagnosedmetabolic disorder allows to establish the optimal dose in a short time(e.g. 7-14 days).

The invention shows major improvement and optimized treatment ofhyperglycaemia and thus allows the possibility of preventing or delayingthe progression or inducing a remission of hyperglycaemia associatedcomplications, in particular diabetic cataract formation, in canineanimals.

A further advantage of the present invention is that the use of one ormore SGLT2 inhibitors, preferably compound A, is effective against themetabolic disorders alone, i.e., if desired the use of one or more SGLT2inhibitors, preferably compound A, in a canine animal provides amonotherapy, i.e. a stand-alone therapy; i.e., no other medication isadministered to the canine animal for the treatment or prevention of thesame metabolic disorder—with the only exemption of insulin dependentdiabetes.

However, the invention also allows for the possibility of combinationtherapy with insulin. Such a combination advantageously leads to adecrease in the dose and/or frequency at which the insulin isadministered, compared to monotherapy of the canine animal with insulin.

Advantageously, the use of one or more SGLT2 inhibitors, preferablycompound A, according to the present invention does not causehypoglycacmia (Example 2).

A further advantage in particular is that the use of one or more SGLT2inhibitors, preferably compound A, according to the present inventionleads to a reduction in insulin resistance in treated, insulin resistantcanine animals. That is, equivalently, the use of one or more SGLT2inhibitors, preferably compound A, according to the present inventionadvantageously leads to increased insulin sensitivity in treated,insulin resistant canine animals.

Thus, use of one or more SGLT2 inhibitors, preferably compound A,according to the present invention provides improved treatment and/orprevention of metabolic diseases as disclosed herein, includingdiabetes, in canine animals.

The effects of using one or more SGLT2 inhibitors, preferably compoundA, according to the present invention may be relative to the same or acomparable canine animal prior to administration of one or more SGLT2inhibitors, preferably compound A according to the present invention,and/or relative to a comparable canine animal that has not received saidtreatment (e.g. a placebo group). In either case, when a comparison ismade, the comparison may be made after a certain treatment period, e.g.,1, 2, 3, 4, 5, 6 or 7 days; 10 days, 14 days; 1, 2, 3, 4, 5, 6, 7 or 8weeks; 1, 2, 3 or 4 months. Preferably the treatment period is 4 weeks.Alternatively, the treatment period may be 6 or 8 weeks. Alternatively,the treatment period may be 8 weeks or more, e.g. 8-16 weeks, i.e. 8, 9,10, 11, 12, 13, 14, 15 or 16 weeks.

A further advantage of the present invention is that one or more SGLT2inhibitors, preferably compound A, may effectively be administered to acanine animal orally. Moreover, the one or more SGLT2 inhibitors,preferably compound A, according to the present invention can beadministered only once per day. These advantages allow for bettercompliance of the treated canine animal and the owner. This leads tobetter glycaemic control of disorders (e.g. diabetes) for which canineanimals are currently treated with insulin. Generally, the use of one ormore SGLT2 inhibitors, preferably compound A, according to the presentinvention thus helps to attenuate (i.e. delays or prevents) theprogression of metabolic disorders and delays or prevents the onset ofmetabolic disorders (e.g. diabetes) and their complications in canineanimals.

The effects of using one or more SGLT2 inhibitors, preferably compoundA, according to the present invention (e.g. the above-mentionedbeneficial effects upon hyperglycaemia) may be relative to the same or acomparable canine animal prior to administration of the one or moreSGLT2 inhibitors, preferably compound A, according to the presentinvention, and/or relative to a comparable canine animal that hasreceived e.g. standard insulin treatment (e.g. a control group) or hasbeen untreated.

A further advantage of the present invention is that the one or moreSGLT2 inhibitors, preferably compound A, may effectively be administeredto a canine animal orally, e.g. in liquid form. Moreover, the one ormore SGLT2 inhibitors, preferably compound A, according to the presentinvention can be administered only once per day. These advantages allowfor optimal dosing and compliance of the treated canine animal andowner.

Accordingly, the present invention also provides pharmaceuticalcompositions comprising one or more SGLT2 inhibitors, preferablycompound A, according to the invention for use in treating and/orpreventing metabolic disorders in canine animals.

The invention also provides methods of treating and/or preventingmetabolic disorders in canine animals, comprising administering to acanine animal in need of such treatment and/or prevention an effectivedose of one or more SGLT2 inhibitors, preferably compound A, asdescribed herein.

Generally, the use of one or more SGLT2 inhibitors, preferably compoundA, according to the present invention may thus attenuate, delay orprevent the progression of a metabolic disorder, e.g. the metabolicdisorders disclosed herein, or may delay the progression or prevent theonset of metabolic disorders and their complications in canine animals,e.g. hypertension, renal dysfunction and/or musculoskeletal disorders isprevented or progression is slowed or remission is achieved.

Definitions

All values and concentrations presented herein are subject to inherentvariations acceptable in biological science within an error of ±10%. Theterm “about” also refers to this acceptable variation.

Treatment effects disclosed herein (such as an improvement, reduction ordelayed onset of a disorder, disease or condition, or the improvement,reduction, increase or delay of any effect, index, marker level or otherparameter relating to a disorder, disease or condition) may be observedwith a statistical significance of p<0.05, preferably <0.01.

When reference is made herein to a deviation (e.g. an increase,elevation, excess, prolongation, raise, reduction, decrease,improvement, delay, abnormal levels, or any other change, alteration ordeviation with respect to a reference), the deviation may be, e.g., by5% or more, particularly 10% or more, more particularly 15% or more,more particularly 20% or more, more particularly 30% or more, moreparticularly 40% or more, or more particularly 50% or more, with respectto the relevant reference value, unless otherwise stated. Typically, thedeviation will be by at least 10%, i.e. 10% or more. The deviation mayalso be by 20%. The deviation may also be by 30%. The deviation may alsobe by 40%. The relevant reference value may be generated from a group ofreference animals which are treated with placebo instead of the one ormore SGLT2 inhibitors, preferably compound A, or are untreated.

Herein, an excursion, e.g. an insulin excursions or glucose excursion,designates a change in concentration or level in blood over time. Themagnitude of excursions, e.g. insulin excursions or glucose excursionsmay be expressed as area-under-curve (AUC) values.

Herein, the terms “active substance” or “active ingredient” encompassone or more SGLT2 inhibitors, preferably compound A, or anypharmaceutically acceptable form thereof (e.g. a prodrug or acrystalline form), for use according to the invention. In the case of acombination with one or additional active compound, the terms “activeingredient” or “active substance” may also include the additional activecompound.

Herein, the expression “clinical condition(s)” refers to pathologiccondition(s) or pathophysiological or physiological changes that arerecognizable, e.g. visible and/or measurable, such as blood parameters,and that are associated with and/or define a disorder and/or disease.

Herein, the expression “associated with”, in particular encompasses theexpression “caused by”.

Herein, ivGTT refers to an intravenous glucose tolerance test. In anivGTT, 0.8 g dextrose per kg body mass may typically be employed.

Herein, ivITT refers to an intravenous insulin tolerance test. In anivITT, 0.05 U insulin per kg body mass may typically be employed.

SGLT2 Inhibitors

SGLT2 inhibitors for use according to the invention include, but are notlimited to, glucopyranosyl-substituted benzene derivatives, for exampleas described in WO 01/27128, WO 03/099836, WO 2005/092877, WO2006/034489, WO 2006/064033, WO 2006/117359, WO 2006/117360, WO2007/025943, WO 2007/028814, WO 2007/031548, WO 2007/093610, WO2007/128749, WO 2008/049923, WO 2008/055870, WO 2008/055940, WO2009/022020 or WO 2009/022008.

Moreover, the one or more SGLT2 inhibitors for use according to theinvention may be selected from the group consisting of the followingcompounds or pharmaceutically acceptable forms thereof:

-   -   (1) a glucopyranosyl-substituted benzene derivative of the        formula (1)

-   -   -   wherein R¹ denotes cyano, Cl or methyl (most preferably            cyano);        -   R² denotes H, methyl, methoxy or hydroxy (most preferably H)            and        -   R³ denotes cyclopropyl, hydrogen, fluorine, chlorine,            bromine, iodine, methyl, ethyl, propyl, isopropyl, butyl,            sec-butyl, iso-butyl, tert-butyl, 3-methyl-but-1-yl,            cyclobutyl, cyclopentyl, cyclohexyl, 1-hydroxy-cyclopropyl,            1-hydroxy-cyclobutyl, 1-hydroxy-cyclopentyl,            1-hydroxy-cyclohexyl, ethinyl, ethoxy, difluoromethyl,            trifluoromethyl, pentafluoroethyl, 2-hydroxyl-ethyl,            hydroxymethyl, 3-hydroxy-propyl,            2-hydroxy-2-methyl-prop-1-yl, 3-hydroxy-3-methyl-but-1-yl,            1-hydroxy-1-methyl-ethyl,            2,2,2-trifluoro-1-hydroxy-1-methyl-ethyl,            2,2,2-trifluoro-1-hydroxy-1-trifluoromethyl-ethyl,            2-methoxy-ethyl, 2-ethoxy-ethyl, hydroxy, difluoromethyloxy,            trifluoromethyloxy, 2-methyloxy-ethyloxy, methylsulfanyl,            methylsulfinyl, methlysulfonyl, ethylsulfinyl,            ethylsulfonyl, trimethylsilyl, (R)-tetrahydrofuran-3-yloxy            or (S)-tetrahydrofuran-3-yloxy or cyano;        -   wherein R3 is preferably selected from cyclopropyl, ethyl,            ethinyl, ethoxy, (R)-tetrahydrofuran-3-yloxy or            (S)-tetrahydrofuran-3-yloxy; and most preferably R3 is            cyclopropyl,        -   or a derivative thereof wherein one or more hydroxyl groups            of the β-D-glucopyranosyl group are acylated with groups            selected from (C₁₋₁₈-alkyl)carbonyl,            (C₁₋₁₈-alkyl)oxycarbonyl, phenylcarbonyl and            phenyl-(C₁₋₃-alkyl)-carbonyl;

    -   (2)        1-cyano-2-(4-cyclopropyl-benzyl)-4-(β-D-glucopyranos-1-yl)-benzene,        represented by formula (2):

-   -   (3) Dapagliflozin, represented by formula (3):

-   -   (4) Canagliflozin, represented by formula (4):

-   -   (5) Empagliflozin, represented by formula (5):

-   -   (6) Luseogliflozin, represented by formula (6):

-   -   (7) Tofogliflozin, represented by formula (7):

-   -   (8) Ipragliflozin, represented by formula (8):

-   -   (9) Ertugliflozin, represented by formula (9):

-   -   (10) Atigliflozin, represented by formula (10):

-   -   (11) Remogliflozin, represented by formula (11):

-   -   (12) a thiophene derivative of the formula (12)

-   -   -   wherein R denotes methoxy or trifluoromethoxy;

    -   (13)        1-(β-D-glucopyranosyl)-4-methyl-3-[5-(4-fluorophenyl)-2-thienylmethyl]benzene        as described in WO 2005/012326, represented by formula (13);

-   -   (14) a spiroketal derivative of the formula (14):

-   -   -   wherein R denotes methoxy, trifluoromethoxy, ethoxy, ethyl,            isopropyl or tert. butyl;

    -   (15) a pyrazole-O-glucoside derivative of the formula (15)

-   -   -   wherein        -   R¹ denotes C₁₋₃-alkoxy,        -   L¹, L² independently of each other denote H or F,        -   R⁶ denotes H, (C₁₋₃-alkyl)carbonyl, (C₁₋₆-alkyl)oxycarbonyl,            phenyloxycarbonyl, benzyloxycarbonyl or benzylcarbonyl;

    -   (16) a compound of the formula (16):

-   -   (17) and Sergliflozin, represented by formula (17):

The term “dapagliflozin” as employed herein refers to dapagliflozin ofthe above structure as well as pharmaceutically acceptable formsthereof, including hydrates and solvates thereof, and crystalline formsthereof. The compound and methods of its synthesis are described in WO03/099836 for example. Preferred hydrates, solvates and crystallineforms are described in the patent applications WO 2008/116179 and WO2008/002824 for example.

The term “canagliflozin” as employed herein refers to canagliflozin ofthe above structure as well as pharmaceutically acceptable formsthereof, including hydrates and solvates thereof, and crystalline formsthereof. The compound and methods of its synthesis are described in WO2005/012326 and WO 2009/035969 for example. Preferred hydrates, solvatesand crystalline forms are described in the patent application WO2008/069327 for example.

The term “empagliflozin” as employed herein refers to empagliflozin ofthe above structure as well as pharmaceutically acceptable formsthereof, including hydrates and solvates thereof, and crystalline formsthereof. The compound and methods of its synthesis are described in WO2005/092877, WO 2006/120208 and WO 2011/039108 for example. A preferredcrystalline form is described in the patent applications WO 2006/117359and WO 2011/039107 for example.

The term “atigliflozin” as employed herein refers to atigliflozin of theabove structure as well as pharmaceutically acceptable forms thereof,including hydrates and solvates thereof, and crystalline forms thereof.The compound and methods of its synthesis are described in WO2004/007517 for example.

The term “ipragliflozin” as employed herein refers to ipragliflozin ofthe above structure as well as pharmaceutically acceptable formsthereof, including hydrates and solvates thereof, and crystalline formsthereof. The compound and methods of its synthesis are described in WO2004/080990, WO 2005/012326 and WO 2007/114475 for example.

The term “tofogliflozin” as employed herein refers to tofogliflozin ofthe above structure as well as pharmaceutically acceptable formsthereof, including hydrates and solvates thereof, and crystalline formsthereof. The compound and methods of its synthesis are described in WO2007/140191 and WO 2008/013280 for example.

The term “luseogliflozin” as employed herein refers to luseogliflozin ofthe above structure as well as pharmaceutically acceptable formsthereof, including hydrates and solvates thereof, and crystalline formsthereof.

The term “ertugliflozin” as employed herein refers to ertugliflozin ofthe above structure as well as pharmaceutically acceptable formsthereof, including hydrates and solvates thereof, and crystalline formsthereof. The compound is described for example in WO 2010/023594.

The term “remogliflozin” as employed herein refers to remogliflozin ofthe above structure as well as pharmaceutically acceptable formsthereof, including prodrugs of remogliflozin, in particularremogliflozin etabonate, including hydrates and solvates thereof, andcrystalline forms thereof. Methods of its synthesis are described in thepatent applications EP 1 213 296 and EP 1 354 888 for example.

The term “sergliflozin” as employed herein refers to sergliflozin of theabove structure as well as pharmaceutically acceptable forms thereof,including prodrugs of sergliflozin, in particular sergliflozinetabonate, including hydrates and solvates thereof, and crystallineforms thereof. Methods for its manufacture are described in the patentapplications EP 1 344 780 and EP 1 489 089 for example.

The compound of formula (16) above and its manufacture are described forexample in WO 2008/042688 or WO 2009/014970.

Preferred SGLT2 inhibitors are glucopyranosyl-substituted benzenederivatives. Optionally, one or more hydroxyl groups of theglucopyranosyl group in such one or more SGLT2 inhibitors may beacylated with groups selected from (C₁₋₁₈-alkyl)carbonyl,(C₁₋₁₈-alkyl)oxycarbonyl, phenylcarbonyl andphenyl-(C₁₋₃-alkyl)-carbonyl.

More preferred are glucopyranosyl-substituted benzonitrile derivativesof formula (1) as disclosed herein above. Yet more preferred areglucopyranosyl-substituted benzonitrile derivatives of formula (18):

whereinR3 denotes cyclopropyl, hydrogen, fluorine, chlorine, bromine, iodine,methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, iso-butyl,tert-butyl, 3-methyl-but-1-yl, cyclobutyl, cyclopentyl, cyclohexyl,1-hydroxy-cyclopropyl, 1-hydroxy-cyclobutyl, 1-hydroxy-cyclopentyl,1-hydroxy-cyclohexyl, ethinyl, ethoxy, difluoromethyl, trifluoromethyl,pentafluoroethyl, 2-hydroxyl-ethyl, hydroxymethyl, 3-hydroxy-propyl,2-hydroxy-2-methyl-prop-1-yl, 3-hydroxy-3-methyl-but-1-yl,1-hydroxy-1-methyl-ethyl, 2,2,2-trifluoro-1-hydroxy-1-methyl-ethyl,2,2,2-trifluoro-1-hydroxy-1-trifluoromethyl-ethyl, 2-methoxy-ethyl,2-ethoxy-ethyl, hydroxy, difluoromethyloxy, trifluoromethyloxy,2-methyloxy-ethyloxy, methylsulfanyl, methylsulfinyl, methlysulfonyl,ethylsulfinyl, ethylsulfonyl, trimethylsilyl,(R)-tetrahydrofuran-3-yloxy or (S)-tetrahydrofuran-3-yloxy or cyano(wherein R3 is preferably selected from cyclopropyl, ethyl, ethinyl,ethoxy, (R)-tetrahydrofuran-3-yloxy or (S)-tetrahydrofuran-3-yloxy; andR3 most preferably is cyclopropyl,or a derivative thereof wherein one or more hydroxyl groups of theβ-D-glucopyranosyl group are acylated with groups selected from(C₁₋₁₈-alkyl)carbonyl, (C₁₋₁₈-alkyl)oxycarbonyl, phenylcarbonyl andphenyl-(C₁₋₃-alkyl)-carbonyl.

Preferably, such SGLT2 inhibitor is1-cyano-2-(4-cyclopropyl-benzyl)-4-(β-D-glucopyranos-1-yl)-benzene asshown in formula (2) (also referred to herein as “compound A”).Optionally, one or more hydroxyl groups of the β-D-glucopyranosyl groupof compound A may be acylated with groups selected from(C₁₋₁₈-alkyl)carbonyl, (C₁₋₁₈-alkyl)oxycarbonyl, phenylcarbonyl andphenyl-(C₁₋₃-alkyl)-carbonyl.

Thus, in preferred embodiments, a SGLT2 inhibitor according to thepresent invention is a glucopyranosyl-substituted benzene derivativeSGLT2 inhibitor, preferably a SGLT2 inhibitor of formula (1), morepreferably of formula (18), or yet more preferably of formula (2) (i.e.compound A), in each case as defined herein above.

Metabolic Disorders

The metabolic disorder may be diabetes, pre-diabetes, obesity and/or anydisorder, disease, condition or symptom associated with one or more ofthose disorders. In particular, the metabolic disorder may behyperglycaemia, impaired glucose tolerance, insulin resistance, insulindependent diabetes and/or hepatic lipidosis. Further relevant metabolicdisorders include hyperinsulinaemia, impaired glucose tolerance, ketosis(in particular ketoacidosis), hyperlipidaemia, dyslipidemia, elevatedblood levels of fatty acids and/or of glycerol, Syndrome X (metabolicsyndrome), and/or inflammation of the pancreas, low grade systemicinflammation, inflammation of adipose tissue.

In some embodiments, the metabolic disorder is diabetes. Herein,diabetes may be pre-diabetes, insulin dependent diabetes or insulinresistance diabetes. In particular, diabetes may be insulin dependentdiabetes.

In some embodiments, the metabolic disorder is hyperglycaemia. Herein,hyperglycaemia may be associated with diabetes, e.g. with insulindependent diabetes or insulin resistance diabetes. In some embodiments,hyperglycaemia may be associated with obesity. The hyperglycaemia may bechronic.

In some embodiments, the metabolic disorder is insulin resistance.Herein, insulin resistance may be associated with diabetes, e.g. withinsulin resistance diabetes. In some embodiments, insulin resistance maybe associated with obesity.

In some embodiments, the metabolic disorder is impaired glucosetolerance (IGT). Herein, impaired glucose tolerance may be associatedwith diabetes, e.g. with insulin dependent diabetes or insulinresistance diabetes. In some embodiments, impaired glucose tolerance maybe associated with obesity.

In some embodiments, the metabolic disorder is hyperinsulinaemia.Herein, hyperinsulinaemia may be associated with diabetes, e.g. withinsulin resistance diabetes. In some embodiments, hyperinsulinaemia maybe associated with obesity.

In some embodiments, the metabolic disorder is one or more ofhyperglycaemia, insulin resistance, and hepatic lipidosis. In someembodiments, the metabolic disorder is selected from hyperglycaemia andinsulin resistance.

In some embodiments, the metabolic disorder is one or more ofhyperinsulinaemia, impaired glucose tolerance, hyperglycaemia andinsulin resistance.

In certain embodiments, the canine animal is obese. For example,according to the invention, one or more metabolic disorders selectedfrom hyperglycaemia, insulin resistance and hepatic lipidosis may betreated and/or prevented in an obese canine animal. Moreover, e.g.,hyperinsulinaemia and/or impaired glucose tolerance may be treatedand/or prevented in an obese canine animal. Moreover, one or moredisorders selected from ketosis (in particular ketoacidosis),hyperlipidaemia, elevated blood levels of fatty acids and/or ofglycerol, Syndrome X (metabolic syndrome), inflammation of the pancreas,inflammation of adipose tissue, may be treated and/or prevented in anobese canine animal.

In certain embodiments, the canine animal is not obese. The metabolicdisorder may be associated and/or caused by e.g. hypo- orhyperthyroidism, hypercortisolism (hyperadrenocorticism, cushing) and/orgrowth-hormone access (acromegaly). For example, according to theinvention, one or more metabolic disorders selected from hyperglycaemia,insulin resistance and hepatic lipidosis may be treated and/or preventedin a non-obese canine animal. Moreover, e.g., hyperinsulinaemia and/orimpaired glucose tolerance may be treated and/or prevented in anon-obese canine animal. Moreover, one or more disorders selected fromketosis (in particular ketoacidosis), hyperlipidaemia, elevated bloodlevels of fatty acids and/or of glycerol, Syndrome X (metabolicsyndrome), inflammation of the pancreas and/or inflammation of adiposetissue may be treated and/or prevented in a non-obese canine animal.

In certain embodiments, the canine animal is suffering from diabetes,e.g. from insulin dependent diabetes or insulin resistance diabetes. Forexample, according to the invention, one or more metabolic disordersselected from the group of hyperglycaemia, impaired glucose toleranceand hepatic lipidosis may be treated and/or prevented in a canine animalthat is suffering from diabetes, e.g. from insulin dependent diabetes orinsulin resistance diabetes. Moreover, e.g., hyperinsulinaemia and/orinsulin resistance may be treated and/or prevented in a canine animalthat is suffering from diabetes, e.g. from or insulin resistancediabetes. Moreover, one or more disorders selected from ketosis (inparticular ketoacidosis), hyperlipidaemia, elevated blood levels offatty acids and/or of glycerol, Syndrome X (metabolic syndrome),inflammation of the pancreas, inflammation of adipose tissue may betreated and/or prevented in a canine animal that is suffering fromdiabetes, e.g. from insulin dependent diabetes or insulin resistancediabetes.

In some embodiments, the canine animal is obese and not suffering fromdiabetes.

In some embodiments, the canine animal is not obese and suffering fromdiabetes.

The present invention also provides the use of one or more SGLT2inhibitors, preferably compound A, for treating and/or preventinghyperglycaemia associated complications. For example by improving thediurnal glycaemic control and thereby delay or prevent the developmentor the progression or induce the regression of cataract formation in acanine animal.

Ketosis is a state of elevated levels of ketone bodies in the body.Ketoacidosis can be described as a type of metabolic acidosis which iscaused by high concentrations of ketone bodies, formed by the breakdownof fatty acids and the deamination of amino acids. The two commonketones produced in humans are acetoacetic acid and β-hydroxybutyrate.In dogs, predominantly three ketones are found: acetoacetic acid,beta-hydroxybutyrate and pyruvic acid. Ketoacidosis can be smelled on asubject's breath. This is due to acetone, a direct byproduct of thespontaneous decomposition of acetoacetic acid.

Ketoacidosis is an extreme and uncontrolled form of ketosis. Ketosis isalso a normal response to prolonged fasting. In ketoacidosis, the bodyfails to adequately regulate ketone production, esp. by producingAcetyl-CoA, causing such a severe accumulation of keto acids that the pHof the blood is substantially decreased, i.e. the excess ketone bodiesmay significantly acidify the blood. In extreme cases ketoacidosis canbe fatal.

Ketoacidosis may occur when the body is producing high levels of ketonebodies via the metabolism of fatty acids (ketosis) and insulin does notsufficiently slow this production (e.g. due to insulinresistance/reduced insulin sensitivity or lack of insulin). The presenceof high blood sugar levels (hyperglycaemia) caused by the lack ofinsulin can lead to further acidity in the blood. In healthy individualsthis normally does not occur because the pancreas produces insulin inresponse to rising ketone/blood sugar levels.

Ketoacidosis is most common in untreated diabetes mellitus, when theliver breaks down fat and proteins in response to a perceived need forrespiratory substrate.

Pre-diabetes in canine animals is characterized by hyperinsulinemia,insulin resistance in target organs, impaired glucose tolerance incl.e.g. an altered insulin response to a glycaemic challenge, e.g. alsoe.g. induced by stress. Pre-diabetes is also often associated withobesity. Pre-diabetes may also be associated with intermittenthyperglycaemia.

Insulin resistance diabetes in canine animals is characterized by bothincreased insulin production and insulin resistance in target organs andas a consequence hyperglycaemia. It is frequently detected in intactfemale diabetic canines and mainly attributed to progesterone acting asan endogenous insulin antagonist. Therefore, it is mostly eitherassociated with the menstrual cycle, i.e. the dioestrus or topregnancy—gestational. Genetic factors, glucosteroids, lack of exercise,and obesity are possible further reasons for insulin resistance.

Clinical signs of diabetes mellitus observed with canine animals includepolydipsia, polyuria, weight loss, and/or polyphagia. In contrast, inother species such as cats anorexia is more often described thanpolyphagia.

Further particularly relevant clinical signs of diabetes mellitus incanine animals within the context of the present invention arehyperglycaemia and glycosuria. Hyperglycaemia in a canine animal (e.g. adog) is defined as plasma glucose values above normal values (3.5-7mmol/l or 60-120 mg/dl), e.g. 8 mmol/l or more or 150 mg/dl or moreplasma glucose. Glycosuria in a canine animal (e.g. a dog) is defined asglucose levels in urine above normal values (0˜2 mmol/L, or 36 mg/dl).The renal threshold is reached with blood glucose concentrations ofapproximately 8-11 mmol/l or 150 to 200 mg/dl.

The diagnosis of diabetes mellitus in canine animals may alternativelybe based on three criteria, e.g., as follows:

-   (1) Fasting blood glucose concentration measurements >250 mg/dl;-   (2) Glycosuria as defined above; and-   (3) One or more of the following: polyuria, polydipsia, polyphagia,    weight loss despite good appetite, or ketonuria (without signs of    severe ketoacidosis).

In addition to the above mentioned diagnostics and in order to supportthem, further examinations can include haematology, blood chemistry,x-ray and/or abdominal ultrasound.

Preferably, the use of the one or more SGLT2 inhibitors, preferablycompound A, according to the invention allows normal or near-normalblood glucose concentrations to be maintained and/or established.However,—unlike for human therapy—this not believed to be alwaysnecessary for diabetic animals and therefore not always the goal of atreatment according to the invention. According to the invention, bloodglucose concentrations may also be maintained, e.g., between 5.5 and16.6 mmol/l or 100 to 300 mg/dl. For canine animals this will often besatisfactory.

Hyperglycaemia induces cataracts are generally acute in onset, rapidlyprogressive, and bilaterally symmetrical. The clouding of the lensinside the eye leads to a decrease or ultimately loss of vision.

Insulin resistance can be described as the condition in which normalamounts of insulin are inadequate to produce a normal insulin responsefrom fat, muscle and liver cells. Insulin resistance in fat cellsreduces the effects of insulin and results in elevated hydrolysis ofstored triglycerides in the absence of measures which either increaseinsulin sensitivity or which provide additional insulin. Increasedmobilization of stored lipids in these cells elevates free fatty acidsin the blood plasma. Insulin resistance in muscle cells reduces glucoseuptake (and so local storage of glucose as glycogen), whereas insulinresistance in liver cells results in impaired glycogen synthesis and afailure to suppress glucose production. Elevated blood fatty acidlevels, reduced muscle glucose uptake, and increased liver glucoseproduction, may all contribute to elevated blood glucose levels(hyperglycaemia). In obese dogs insulin resistance, i.e. a 5-fold lowerinsulin sensitivity than in lean dogs is detected.

Insulin resistance may be present in association with obesity, visceraladiposity, hypertension and dyslipidaemia involving elevatedtriglycerides, small dense low-density lipoprotein (sdLDL) particles,and decreased HDL cholesterol levels. With respect to visceraladiposity, a great deal of evidence in humans suggests two strong linkswith insulin resistance. First, unlike subcutaneous adipose tissue,visceral adipose cells produce significant amounts of proinflammatorycytokines such as tumour necrosis factor-alpha (TNF-alpha), andInterleukins-1 and -6, etc. In numerous experimental models, theseproinflammatory cytokines profoundly disrupt normal insulin action infat and muscle cells, and may be a major factor in causing thewhole-body insulin resistance observed in human patients with visceraladiposity. Similarly, in canines excessive fat depots contribute to lowgrade systemic inflammation. The cause of the vast majority of cases ofinsulin resistance remains unknown. There is clearly an inheritedcomponent. However, there are some grounds for suspecting that insulinresistance is related to a high-carbohydrate diet. Inflammation alsoseems to be implicated in causing insulin resistance.

Hyperinsulinaemia can be described as a condition in which there areexcess levels, i.e. more than about 35 pmol/L under basal or about 200pmol/L during e.g. a glycaemic challenge (e.g. ivGTT or stress) ofinsulin circulating in the blood. As mentioned, it is commonly presentin cases of, and may be a consequence of, insulin resistance in canineanimals.

Impaired glucose tolerance can be described as condition in which theresponse to a after a glycaemic challenge e.g. after a meal or after aloading test (glucose tolerance test) or after stress induced elevationof blood glucose concentration, the glycaemic peak of the glucoseexcursion is higher and/or the duration of the glucose excursion isprolonged.

Dyslipidaemia or hyperlipidaemia is the presence of raised or abnormallevels of lipids and/or lipoproteins in the blood. Lipid and lipoproteinabnormalities are regarded as a highly modifiable risk factor forcardiovascular disease due to the influence of cholesterol. Glycerol isa precursor for the synthesis of triacylglycerols (triglycerids) and ofphospholipids in the liver and adipose tissue. When the body uses storedfat as a source of energy, glycerol and fatty acids are released intothe bloodstream after hydrolysis of the triglycerides. The glycerolcomponent can be converted to glucose by the liver and provides energyfor cellular metabolism. Normal levels of free fatty acids in the bloodof companion (such as canine) animals are triglyceride concentrations of50 to 150 mg/dl. Normal levels of blood cholesterol are, e.g., 130 to300 mg/dl for the dog.

Dysadipokinemia can be described as a condition in which the circulatingplasma levels of biologically active substances produced in adiposetissue that act in an autocrine/paracrine or endocrine fashion isdeviated, e.g. an elevation of leptin and/or a reduction of adiponectin.

Subclinical inflammation or systemic inflammation, in particular lowgrade systemic inflammation is characterized by increased expression andsecretion of pro-inflammatory cytokines such as tumour necrosisfactor-alpha and/or lower expression and secretion of anti-inflammatorycytokines e.g. interleukin-10 and/or their respective receptors.

Obesity can be described as a medical condition in which excess body fathas accumulated to the extent that it may have an adverse effect onhealth, leading to reduced life expectancy. In obese canine, e.g. a bodycondition score (BCS) of larger than 7 (out of 9) is encountered.

Metabolic disorders to be treated and/or prevented according to theinvention include Syndrome X (metabolic syndrome). This disorder can bedescribed as a combination of medical disorders that increase the riskof developing manifest clinical consequences—e.g. hypertension,cardiomyopathy, renal dysfunction and/or musculoskeletal disorders incanine animals.

Metabolic syndrome is also known as metabolic Syndrome X (metabolicsyndrome), Syndrome X (metabolic syndrome), insulin resistance syndrome,Reaven's syndrome, and CHAOS (as an abbreviation for Coronary arterydisease, Hypertension, Atherosclerosis, Obesity, and Stroke).

The exact mechanisms of the complex pathways of metabolic syndrome arenot yet completely known. The pathophysiology is extremely complex andhas been only partially elucidated. Most patients are older, obese,sedentary, and have a degree of insulin resistance. The most importantfactors in order are: (1) overweight and obesity, (2) genetics, (3)aging, and (4) sedentary lifestyle, i.e., low physical activity andexcess caloric intake.

The pathophysiology is commonly characterized by the development ofvisceral fat after which the adipocytes (fat cells) of the visceral fatincrease plasma levels of TNF-alpha and alter levels of a number ofother substances (e.g., adiponectin, resistin, PAI-1). TNF-alpha hasbeen shown not only to cause the production of inflammatory cytokines,but possibly to trigger cell signalling by interaction with a TNF-alphareceptor that may lead to insulin resistance.

Current first line treatment is change of lifestyle (i.e., caloricrestriction and physical activity). However, drug treatment isfrequently required. Accordingly, the present invention also providesfor prevention of clinically relevant consequences of the metabolicdisorder e.g. hypertension, cardiomyopathy, renal dysfunction and formusculoskeletal disorders in canine animals.

Metabolic disorders to be treated and/or prevented according to theinvention include inflammation of the pancreas (pancreatitis). Thisdisorder may occur as either an acute form or a chronic form. Chronicpancreatitis may occur with or without steatorrhea and/or diabetesmellitus.

Pancreatitis may be caused by hypertriglyceridemia (in particular whentriglyceride values exceed 1500 mg/dl (16 mmol/l), hypercalcemia, viralinfection, trauma, vasculitis (i.e. inflammation of the small bloodvessels within the pancreas), and autoimmune pancreatitis.

Metabolic disorders, esp. dyslipidaemia and elevated serum levels oftriglycerides are risk factors for the development of pancreatitis, andmay thus be treated according to the present invention in associationwith pancreatitis. Accordingly, the present invention also provides forprevention of pancreatitis.

Metabolic disorders to be treated and/or prevented according to theinvention include an inflammation of adipose tissue (panniculitis),which is a group of disorders characterised by inflammation ofsubcutaneous adipose tissue.

Panniculitis may occur in any fatty tissue (cutaneous and/or visceral).It may be diagnosed on the basis of a deep skin biopsy, and can befurther classified by histological characteristics based on the locationof the inflammatory cells (within fatty lobules or in the septa whichseparate them) and on the presence or absence of vasculitis.Panniculitis can also be classified based on the presence or absence ofsystemic symptoms.

Metabolic diseases, esp. pancreatitis, are risk factors for thedevelopment of panniculitis, and may thus be treated according to thepresent invention in association with panniculitis. Accordingly, thepresent invention also provides for prevention of panniculitis.

Canine animals Herein, a canine animal may be a member of the Canidaefamily (i.e. a canid). It may thus belong either to the subfamily Canini(related to wolves) or Vulpini (related to foxes). The term canineanimal encompasses the term dog, e.g., a domestic dog. The term domesticdog encompasses the terms Canis lupus familiaris and Canis lupus dingo.

Pharmaceutically Acceptable Forms

Herein, references to SGLT2 inhibitors and/or their use according to theinvention encompass pharmaceutically acceptable forms of the SGLT2inhibitors, unless otherwise stated.

According to the invention, any pharmaceutically acceptable form of theSGLT2 inhibitor, e.g. of formula (1), preferably formula (18), morepreferably formula (2), may be used. E.g. a crystalline form may beused. Prodrug forms are also encompassed by the present invention.

Prodrug forms may include, e.g., esters and/or hydrates. The termprodrug is also meant to include any covalently bonded carrier whichreleases the active compound of the invention in vivo when the prodrugis administered to a mammalian subject. Prodrugs of a compound of theinvention may be prepared by modifying functional groups present in thecompound of the invention in such a way that the modifications arecleaved, either in routine manipulation or in vivo, to the parentcompound of the invention.

Crystalline forms for use according to the invention include a complexof an SGLT2 inhibitor with one or more amino acids (see e.g. WO2014/016381). An amino acid for such use may be a natural amino acid.The amino acid may be a proteogenic amino acid (includingL-hydroxyproline), or a non-proteogenic amino acid. The amino acid maybe a D- or an L-amino acid. In some preferred embodiments the amino acidis proline (L-proline and/or D-proline, preferably L-proline). E.g., acrystalline complex of1-cyano-2-(4-cyclopropyl-benzyl)-4-(β-D-glucopyranos-1-yl)-benzene(formula (2); compound A) with proline (e.g. L-proline) is preferred.

Thus, herein is disclosed a crystalline complex between one or morenatural amino acids and an SGLT2 inhibitor, e.g., a crystalline complexbetween one or more natural amino acids and a glucopyranosyl-substitutedbenzene derivative SGLT2 inhibitor, preferably a SGLT2 inhibitor offormula (1), more preferably of formula (18) or yet more preferably offormula (2) (compound A). Thus, herein is disclosed a crystallinecomplex between one or more natural amino acids and1-cyano-2-(4-cyclopropyl-benzyl)-4-(β-D-glucopyranos-1-yl)-benzene(compound A).

Further disclosed herein is the use of one or more crystalline complexesas defined hereinbefore or hereinafter for preparing a pharmaceuticalcomposition which is suitable for the treatment or prevention ofdiseases or conditions which can be influenced by inhibitingsodium-dependent glucose co-transporter SGLT, preferably SGLT2. Furtherdisclosed herein is the use of one or more crystalline complexes asdefined hereinbefore or hereinafter for preparing a pharmaceuticalcomposition for inhibiting the sodium-dependent glucose co-transporterSGLT2.

A crystalline complex between one or more natural amino acids (e.g.proline, preferably L-proline) and an SGLT2 inhibitor, is a preferredpharmaceutically acceptable form of a SGLT2 inhibitor for use accordingto the present invention. In particular, a crystalline complex betweenone or more natural amino acids (e.g. proline, preferably L-proline) anda glucopyranosyl-substituted benzene derivative SGLT2 inhibitor,preferably a SGLT2 inhibitor of formula (1), more preferably of formula(18) or yet more preferably of formula (2) (compound A) is a preferredpharmaceutically acceptable form of a SGLT2 inhibitor for use accordingto the present invention. A crystalline complex between one or morenatural amino acids (e.g. proline, preferably L-proline) and1-cyano-2-(4-cyclopropyl-benzyl)-4-(β-D-glucopyranos-1-yl)-benzene(compound A) is particularly preferred as a pharmaceutically acceptableform of a SGLT2 inhibitor for use according to the present invention.

Also disclosed herein is a method for making one or more crystallinecomplexes as defined hereinbefore and hereinafter, said methodcomprising the following steps:

(a) preparing a solution of the SGLT2 inhibitor (e.g. aglucopyranosyl-substituted benzene derivative, or a SGLT2 inhibitor offormula (1), preferably formula (18) or more preferably formula (2),i.e. compound A) and the one or more natural amino acids in a solvent ora mixture of solvents;(b) storing the solution to precipitate the crystalline complex out ofsolution;(c) removing the precipitate from the solution; and(d) drying the precipitate optionally until any excess of said solventor mixture of solvents has been removed.

A certain pharmaceutical activity is of course the basic prerequisite tobe fulfilled by a pharmaceutically active agent before same is approvedas a medicament on the market. However, there are a variety ofadditional requirements a pharmaceutically active agent has to complywith. These requirements are based on various parameters which areconnected with the nature of the active substance itself. Without beingrestrictive, examples of these parameters are the stability of theactive agent under various environmental conditions, its stabilityduring production of the pharmaceutical formulation and the stability ofthe active agent in the final medicament compositions. Thepharmaceutically active substance used for preparing the pharmaceuticalcompositions should be as pure as possible and its stability inlong-term storage must be guaranteed under various environmentalconditions. This is essential to prevent the use of pharmaceuticalcompositions which contain, in addition to the actual active substance,breakdown products thereof, for example. In such cases the content ofactive substance in the medicament might be less than that specified.

Uniform distribution of the medicament in the formulation is a criticalfactor, particularly when the medicament has to be given in low doses.To ensure uniform distribution, the particle size of the activesubstance can be reduced to a suitable level, e.g. by grinding. Sincebreakdown of the pharmaceutically active substance as a side effect ofthe grinding (or micronising) has to be avoided as far as possible, inspite of the hard conditions required during the process, it isessential that the active substance should be highly stable throughoutthe grinding process. Only if the active substance is sufficientlystable during the grinding process it is possible to produce ahomogeneous pharmaceutical formulation which always contains thespecified amount of active substance in a reproducible manner.

Another problem which may arise in the grinding process for preparingthe desired pharmaceutical formulation is the input of energy caused bythis process and the stress on the surface of the crystals. This may incertain circumstances lead to polymorphous changes, to amorphization orto a change in the crystal lattice. Since the pharmaceutical quality ofa pharmaceutical formulation requires that the active substance shouldalways have the same crystalline morphology, the stability andproperties of the crystalline active substance are subject to stringentrequirements from this point of view as well.

The stability of a pharmaceutically active substance is also importantin pharmaceutical compositions for determining the shelf life of theparticular medicament; the shelf life is the length of time during whichthe medicament can be administered without any risk. High stability of amedicament in the abovementioned pharmaceutical compositions undervarious storage conditions is therefore an additional advantage for boththe patient and the manufacturer.

The absorption of moisture reduces the content of pharmaceuticallyactive substance as a result of the increased weight caused by theuptake of water. Pharmaceutical compositions with a tendency to absorbmoisture have to be protected from moisture during storage, e.g. by theaddition of suitable drying agents or by storing the drug in anenvironment where it is protected from moisture. Preferably, therefore,a pharmaceutically active substance should be at best slightlyhygroscopic.

Furthermore, the availability of a well-defined crystalline form allowsthe purification of the drug substance by recrystallization.

Apart from the requirements indicated above, it should be generallyborne in mind that any change to the solid state of a pharmaceuticalcomposition which is capable of improving its physical and chemicalstability gives a significant advantage over less stable forms of thesame medicament.

A crystalline complex between a natural amino acid and an SGLT2inhibitor (e.g. a glucopyranosyl-substituted benzene derivative or aSGLT2 inhibitor of formula (1), or formula (18) or, particularly, offormula (2), i.e. compound A) fulfills important requirements mentionedhereinbefore.

Preferably the natural amino acid is present in either its (D) or (L)enantiomeric form, most preferably as the (L) enantiomer.

Furthermore those crystalline complexes according to this invention arepreferred which are formed between the SGLT2 inhibitor (e.g. of formula(1), preferably formula (18) or, particularly, of formula (2), i.e.compound A) and one natural amino acid, most preferably between thecompound A and the (L) enantiomer of a natural amino acid.

Preferred amino acids according to this invention are selected from thegroup consisting of phenylalanine and proline, in particular (L)-prolineand (L)-phenylalanine.

According to a preferred embodiment the crystalline complex ischaracterized in that the natural amino acid is proline, in particular(L)-proline.

Preferably the molar ratio of the SGLT2 inhibitor (e.g. of formula (1),preferably formula (18) or, particularly, of formula (2), i.e. compoundA) and the natural amino acid is in the range from about 2:1 to about1:3; more preferably from about 1.5:1 to about 1:1.5, even morepreferably from about 1.2:1 to about 1:1.2, most preferably about 1:1.In the following such an embodiment is referred to as “complex (1:1)” or“1:1 complex”.

Therefore a preferred crystalline complex according to this invention isa complex (1:1) between said SGLT2 inhibitor (e.g. of formula (1),preferably formula (18) or, particularly, of formula (2), i.e. compoundA) and proline; in particular of said SGLT2 inhibitor and L-proline.

According to a preferred embodiment the crystalline complex, in theparticular the 1:1 complex of said SGLT2 inhibitor with L-proline, is ahydrate.

Preferably the molar ratio of the crystalline complex and water is inthe range from about 1:0 to 1:3; more preferably from about 1:0 to 1:2,even more preferably from about 1:0.5 to 1:1.5, most preferably about1:0.8 to 1:1.2, in particular about 1:1.

The crystalline complex of said SGLT2 inhibitor with proline, inparticular with L-proline and water, may be identified and distinguishedfrom other crystalline forms by means of their characteristic X-raypowder diffraction (XRPD) patterns.

For example, a crystalline complex of compound A with L-proline ispreferably characterised by an X-ray powder diffraction pattern thatcomprises peaks at 20.28, 21.14 and 21.64 degrees 2Θ (±0.1 degrees 2Θ),wherein said X-ray powder diffraction pattern is made using CuK_(α1)radiation.

In particular said X-ray powder diffraction pattern comprises peaks at4.99, 20.28, 21.14, 21.64 and 23.23 degrees 2Θ (±0.1 degrees 2Θ),wherein said X-ray powder diffraction pattern is made using CuK_(α1)radiation.

More specifically said X-ray powder diffraction pattern comprises peaksat 4.99, 17.61, 17.77, 20.28, 21.14, 21.64, 23.23 and 27.66 degrees 2Θ(±0.1 degrees 2Θ), wherein said X-ray powder diffraction pattern is madeusing CuK_(α1) radiation.

Even more specifically said X-ray powder diffraction pattern comprisespeaks at 4.99, 15.12, 17.61, 17.77, 18.17, 20.28, 21.14, 21.64, 23.23and 27.66 degrees 2Θ (±0.1 degrees 2Θ), wherein said X-ray powderdiffraction pattern is made using CuK_(α1) radiation.

Even more specifically, the crystalline complex of compound A andL-proline is characterised by an X-ray powder diffraction pattern, madeusing CuK_(α1) radiation, which comprises peaks at degrees 2Θ (±0.1degrees 2Θ) as contained in Table 1.

TABLE 1 X-ray powder diffraction pattern of the crystalline complex ofcompound A and L-proline (only peaks up to 30° in 2Θ are listed): 2Θd-value Intensity I/I₀ [°] [Å] [%] 4.99 17.68 39 7.01 12.61 6 8.25 10.7011 9.95 8.88 12 13.15 6.73 30 13.33 6.64 10 14.08 6.28 4 15.12 5.85 3216.40 5.40 12 16.49 5.37 13 17.11 5.18 6 17.61 5.03 32 17.77 4.99 3518.17 4.88 32 18.32 4.84 28 18.72 4.74 8 19.16 4.63 30 19.96 4.45 2620.28 4.37 56 20.60 4.31 7 21.14 4.20 84 21.64 4.10 100 22.33 3.98 1523.23 3.83 41 24.06 3.70 4 24.51 3.63 15 24.93 3.57 26 25.89 3.44 2326.21 3.40 11 26.84 3.32 8 27.66 3.22 38 27.96 3.19 9 28.26 3.16 5 28.443.14 6 28.75 3.10 6 29.18 3.06 19

Even more specifically, said crystalline complex is characterised by anX-ray powder diffraction pattern, made using CuK_(α1) radiation, whichcomprises peaks at degrees 2Θ (±0.1 degrees 2Θ as shown in FIG. 3.Furthermore said crystalline complex of the compound A with L-proline ischaracterized by a melting point of above 89° C., in particular in arange from about 89° C. to about 115° C., more preferably in a rangefrom about 89° C. to about 110° C. (determined via DSC; evaluated asonset-temperature; heating rate 10 K/min). It can be observed that thiscrystalline complex melts under dehydration. The obtained DSC curve isshown in FIG. 4.

Said crystalline complex of the compound A with L-proline shows a weightloss by thermal gravimetry (TG). The observed weight loss indicates thatthe crystalline form contains water which may be bound by adsorptionand/or may be part of the crystalline lattice, i.e. the crystalline formmay be present as a crystalline hydrate. The content of water in thecrystalline form lies in the range from 0 to about 10 weight-%, inparticular 0 to about 5 weight-%, even more preferably from about 1.5 toabout 5 weight-%. The dotted line in FIG. 2 depicts a weight loss ofbetween 2.8 and 3.8% of water. From the observed weight loss astoichiometry close to a monohydrate can be estimated.

Said crystalline complex has advantageous physicochemical propertieswhich are beneficial in the preparation of a pharmaceutical composition.In particular the crystalline complex has a high physical and chemicalstability under various environmental conditions and during theproduction of a medicament. For example the crystals can be obtained ina shape and particle size which are particular suitable in a productionmethod for solid pharmaceutical formulations. In addition the crystalsshow a high mechanical stability that allows grinding of the crystals.Furthermore the crystalline complex does not show a high tendency toabsorb moisture and is chemically stable, i.e. the crystalline complexallows the production of a solid pharmaceutical formulation with a longshelf life. On the other hand the crystalline complex has a favorablyhigh solubility over a wide pH-range which is advantageous in solidpharmaceutical formulations for oral administration.

The X-ray powder diffraction patterns may be recorded using a STOE-STADIP-diffractometer in transmission mode fitted with a location-sensitivedetector (OED) and a Cu-anode as X-ray source (CuK_(α1) radiation,λ=1.54056 Å, 40 kV, 40 mA). In Table 1 the values “2Θ [°]” denote theangle of diffraction in degrees and the values “d [Å]” denote thespecified distances in Å between the lattice planes. The intensity shownin FIG. 3 is given in units of cps (counts per second).

In order to allow for experimental error, the above described 2Θ valuesshould be considered accurate to ±0.1 degrees 2Θ, in particular ±0.05degrees 2Θ. That is to say, when assessing whether a given sample ofcrystals of the compound A is the crystalline form in accordance withthe above described 2Θ values, a 2Θ value which is experimentallyobserved for the sample should be considered identical with acharacteristic value described above if it falls within ±0.1 degrees 2Θof the characteristic value, in particular if it falls within ±0.05degrees 2Θ of the characteristic value.

The melting point is determined by DSC (Differential ScanningCalorimetry) using a DSC 821 (Mettler Toledo). The weight loss isdetermined by thermal gravimetry (TG) using a TGA 851 (Mettler Toledo).

Also disclosed herein is a method for making a crystalline complex asdefined hereinbefore and hereinafter, said method comprising thefollowing steps:

(a) preparing a solution of an SGLT2 inhibitor as described herein (e.g.compound A or another SGLT2 inhibitor described herein) and the one ormore natural amino acids in a solvent or a mixture of solvents;

(b) storing the solution to precipitate the crystalline complex out ofsolution;

(c) removing the precipitate from the solution; and

(d) drying the precipitate optionally until any excess of said solventor mixture of solvents has been removed.

According to step (a) a solution of the SGLT2 inhibitor (e.g. compound Aor another SGLT2 inhibitor described herein) and the one or more naturalamino acids in a solvent or a mixture of solvents is prepared.Preferably the solution is saturated or at least nearly saturated oreven supersaturated with respect to the crystalline complex. In the step(a) the SGLT2 inhibitor may be dissolved in a solution comprising theone or more natural amino acids or the one or more natural amino acidsmay be dissolved in a solution comprising the SGLT2 inhibitor. Accordingto an alternative procedure the SGLT2 inhibitor is dissolved in asolvent or mixture of solvents to yield a first solution and the one ormore natural amino acids are dissolved in a solvent or mixture ofsolvents to yield a second solution. Thereafter said first solution andsaid second solution are combined to form the solution according to step(a).

Preferably the molar ratio of the natural amino acid and the SGLT2inhibitor (e.g. compound A or any other SGLT2 inhibitor describedherein) in the solution corresponds to the molar ratio of the naturalamino acid and the SGLT2 inhibitor in the crystalline complex to beobtained. Therefore a preferred molar ratio is in the range from about1:2 to 3:1; most preferably about 1:1.

Suitable solvents are preferably selected from the group consisting ofC₁₋₄-alkanols, water, ethylacetate, acetonitrile, acetone, diethylether,tetrahydrofuran, and mixture of two or more of these solvents.

More preferred solvents are selected from the group consisting ofmethanol, ethanol, isopropanol, water and mixture of two or more ofthese solvents, in particular mixtures of one or more of said organicsolvents with water.

Particularly preferred solvents are selected from the group consistingof ethanol, isopropanol, water and mixtures of ethanol and/orisopropanol with water.

In case a mixture of water and one or more C₁₋₄-alkanols, in particularof methanol, ethanol and/or isopropanol, most preferably of ethanol, istaken, a preferred volume ratio of water:the alkanol is in the rangefrom about 99:1 to 1:99; more preferably from about 50:1 to 1:80; evenmore preferably from about 10:1 to 1:60.

Preferably the step (a) is carried out at about room temperature (about20° C.) or at an elevated temperature up to about the boiling point ofthe solvent or mixture of solvents used.

According to a preferred embodiment the starting material of the SGLT2inhibitor (e.g. compound A or any other SGLT2 inhibitor describedherein) and/or of the one or more natural amino acids and/or of thesolvent and mixtures of solvents contain an amount of H₂O which is atleast the quantity required to form a hydrate of the SGLT2 inhibitor; inparticular at least 1 mol, preferably at least 1.5 mol of water per molof SGLT2 inhibitor. Even more preferably the amount of water is at least2 mol of water per mol of SGLT2 inhibitor. This means that either theSGLT2 inhibitor (e.g. compound A) as starting material or the one ormore natural amino acids or said solvent or mixture of solvents, or saidcompounds and/or solvents in combination contain an amount of H₂O asspecified above. For example if the starting material of the SGLT2inhibitor (e.g. compound A) or of the natural amino acid in step (a)does contain sufficient water as specified above, a water content of thesolvent(s) is not mandatory.

In order to reduce the solubility of the crystalline complex accordingto this invention in the solution, in step (a) and/or in step (b) one ormore antisolvents may be added, preferably during step (a) or at thebeginning of step (b). Water is an example of a suitable antisolvent.The amount of antisolvent is preferably chosen to obtain asupersaturated or saturated solution with respect to the crystallinecomplex.

In step (b) the solution is stored for a time sufficient to obtain aprecipitate, i.e. the crystalline complex. The temperature of thesolution in step (b) is about the same as in or lower than in step (a).During storage the temperature of the solution is preferably lowered,preferably to a temperature in the range of 20° C. to 0° C. or evenlower. The step (b) can be carried out with or without stirring. Asknown to the one skilled in the art by the period of time and thedifference of temperature in step (b) the size, shape and quality of theobtained crystals can be controlled. Furthermore the crystallization maybe induced by methods as known in the art, for example by mechanicalmeans such as scratching or rubbing the contact surface of the reactionvessel for example with a glass rod. Optionally the (nearly) saturatedor supersaturated solution may be inoculated with seed crystals.

In step (c) the solvent(s) can be removed from the precipitate by knownmethods as for example filtration, suction filtration, decantation orcentrifugation.

In step (d) an excess of the solvent(s) is removed from the precipitateby methods known to the one skilled in the art as for example byreducing the partial pressure of the solvent(s), preferably in vacuum,and/or by heating above ca. 20° C., preferably in a temperature rangebelow 100° C., even more preferably below 85° C.

Compound A may be synthesized by methods as specifically and/orgenerally described or cited in international application WO 2007/128749which in its entirety is incorporated herein by reference, and/or in theExamples disclosed herein below. Biological properties of the compound Amay also be investigated as is described in WO 2007/128749.

A crystalline complex as described herein is preferably employed as drugactive substance in substantially pure form, that is to say, essentiallyfree of other crystalline forms of the SGLT2 inhibitor (e.g. compoundA). Nevertheless, the invention also embraces a crystalline complex inadmixture with another crystalline form or forms. Should the drug activesubstance be a mixture of crystalline forms, it is preferred that thesubstance comprises at least 50%-weight, even more preferably at least90%-weight, most preferably at least 95%-weight of the crystallinecomplex as described herein.

In view of its ability to inhibit SGLT activity, a crystalline complexaccording to the invention is suitable for the use in the treatmentand/or preventive treatment of conditions or diseases which may beaffected by the inhibition of SGLT activity, particularly SGLT-2activity, in particular the metabolic disorders as described herein. Thecrystalline complex according to the invention is also suitable for thepreparation of pharmaceutical compositions for the treatment and/orpreventive treatment of conditions or diseases which may be affected bythe inhibition of SGLT activity, particularly SGLT-2 activity, inparticular metabolic disorders as described herein. A crystallinecomplex as described herein (in particular of compound A with a naturalamino acid, e.g. proline, particularly L-proline) is also suitable forthe use in the treatment of canines.

Pharmaceutical Compositions and Formulations

The one or more SGLTS inhibitors, preferably compound A, for useaccording to the invention may be prepared as pharmaceuticalcompositions. It may be prepared as solid or as liquid formulations. Ineither case, it is preferably prepared for oral administration,preferably in liquid form for oral administration. The one or more SGLTSinhibitors, preferably compound A, may, however, also be prepared, e.g.,for parenteral administration.

Solid formulations include tablets, granular forms, and other solidforms such as suppositories. Among solid formulations, tablets andgranular forms are preferred.

Pharmaceutical compositions within the meaning of the present inventionmay comprise one or more SGLT2 inhibitors, preferably compound A,according to the present invention and one or more excipients. Anyexcipient that allows for, or supports, the intended medical effect maybe used. Such excipients are available to the skilled person. Usefulexcipients are for example antiadherents (used to reduce the adhesionbetween the powder (granules) and the punch faces and thus preventsticking to tablet punches), binders (solution binders or dry bindersthat hold the ingredients together), coatings (to protect tabletingredients from deterioration by moisture in the air and make large orunpleasant-tasting tablets easier to swallow), disintegrants (to allowthe tablet to break upon dilution), fillers, diluents, flavours,colours, glidants (flow regulators—to promote powder flow by reducinginterparticle friction and cohesion), lubricants (to prevent ingredientsfrom clumping together and from sticking to the tablet punches orcapsule filling machine), preservatives, sorbents, sweeteners etc.

Formulations according to the invention, e.g. solid formulations, maycomprise carriers and/or disintegrants selected from the group of sugarsand sugar alcohols, e.g. mannitol, lactose, starch, cellulose,microcrystalline cellulose and cellulose derivatives, e. g.methylcellulose, and the like.

Manufacturing procedures for formulations suitable for canine animalsare known to the person skilled in the art, and for solid formulationscomprise, e.g., direct compression, dry granulation and wet granulation.In the direct compression process, the active ingredient and all otherexcipients are placed together in a compression apparatus that isdirectly applied to press tablets out of this material. The resultingtablets can optionally be coated afterwards in order to protect themphysically and/or chemically, e.g. by a material known from the state ofthe art.

A unit for administration, e.g. a single liquid dose or a unit of asolid formulation, e.g. a tablet, may comprise 0.01 mg to 10 mg, or e.g.0.3 mg to 1 mg, 1 mg to 3 mg, 3 mg to 10 mg; or 5 to 2500 mg, or e.g. 5to 2000 mg, 5 mg to 1500 mg, 10 mg to 1500 mg, 10 mg to 1000 mg, or10-500 mg of an SGLT2 inhibitor for use according to the invention. Asthe skilled person would understand, the content of the SGLT2 inhibitorin a solid formulation, or any formulation as disclosed herein foradministration to a canine animal, may be increased or decreased asappropriate in proportion to the body weight of the canine animal to betreated.

In one embodiment a pharmaceutical composition for use according to theinvention is designed for oral or parenteral administration, preferablyfor oral administration. Especially the oral administration isameliorated by excipients which modify the smell and/or hapticproperties of the pharmaceutical composition for the intended patient,e.g. as described.

When the SGLT2 inhibitor for use according to the invention isformulated for oral administration, it is preferred that excipientsconfer properties, e.g. palatability and/or chewability that render theformulation suitable for administration to a canine animal.

Also preferred are liquid formulations. Liquid formulations may be,e.g., solutions, syrups or suspensions. They may be administereddirectly to the canine animal or may be mixed with the food and/or drink(e.g. drinking water, or the like) of the canine animal. One advantageof a liquid formulation (similar to a formulation in granular form), isthat such a dosage form allows precise dosing. For example, the SGLT2inhibitor may be dosed precisely in proportion to the body weight of acanine animal. Typical compositions of liquid formulations are known tothe person skilled in the art.

Dosing and Administration

A practitioner skilled in the art can determine suitable doses for theuses of the present invention. Preferred units dosing units includemg/kg, i.e. mg SGLT2 inhibitor per body weight of the canine animal. AnSGLT2 inhibitor of the invention may, e.g., be administered in doses of0.01-5.0 mg/kg body weight per day, e.g. 0.01-4.0 mg/kg body weight perday, e.g. 0.01-3.0 mg/kg body weight per day, e.g. 0.01-2.0 mg/kg bodyweight per day, e.g. 0.01-1.5 mg/kg body weight per day, e.g., 0.01-1.0mg/kg body weight per day, e.g. 0.01-0.75 mg/kg body weight per day,e.g. 0.01-0.5 mg/kg body weight per day, e.g. 0.01-0.4 mg/kg body weightper day, e.g. 0.01-0.3 mg/kg body weight per day, or 0.1 to 3.0 mg/kgbody weight per day, preferably from 0.2 to 2.0 mg/kg body weight perday, more preferably from 0.1 to 1 mg/kg body weight per day. In anotherpreferred embodiment the dose is 0.02-0.5 mg/kg body weight per day,more preferably 0.03-0.4 mg/kg body weight per day, e.g. 0.03-0.3 mg/kgbody weight per day.

A practitioner skilled in the art is able to prepare an SGLT2 inhibitorof the invention for administration according to a desired dose.

Preferably, according to the invention, an SGLT2 inhibitor isadministered no more than three times per day, more preferably no morethan twice per day, most preferably only once per day. The frequency ofadministration can be adapted to the typical feeding rate of the canineanimal.

According to the invention, an SGLT2 inhibitor may be administered suchthat an appropriate blood plasma concentration of the SGLT2 inhibitorsis achieved (e.g. a maximal blood plasma concentration, or blood plasmaconcentration after a given time, e.g. 4, 8, 12 or 24 hours after oraladministration, preferably about 8 hours after oral administration).E.g., for compound A, the blood plasma concentration (e.g. maximal bloodplasma concentration or blood plasma concentration after said given timeafter oral administration) may be within the range 2 to 4000 nM, e.g. 20to 3000 nM, or e.g. 40 to 2000 nM.

Preferably, following administration and the time required for an SGLT2inhibitor to reach the bloodstream, such levels are maintained in theblood over a time interval of at least 12 hours, more preferably atleast 18 hours, most preferably at least 24 h.

Preferably, according to the invention, an SGLT2 inhibitor isadministered orally, in liquid or solid form. The SGLT2 inhibitor may beadministered directly to the animals mouth (e.g. using a syringe,preferably a body-weight-graduated syringe) or together with theanimal's food or drink (e.g. with its drinking water or the like), ineach case preferably in liquid form. The SGLT2 inhibitor may, however,also be administered, e.g., parenterally, or by any other route ofadministration, e.g., rectally.

The SGLT2 inhibitor may be used alone or in combination with anotherdrug. In some embodiments, the one or more SGLTS inhibitors, preferablycompound A, is used in combination with one or more further oralantihyperglycaemic drugs. When the SGLT2 inhibitor is used incombination with a further drug, the SGLT2 inhibitor and any furtherdrug may be administered simultaneously, sequentially (in any order),and/or according to a chronologically staggered dosage regime. In suchembodiments, when a further drug for combined administration with anSGLT2 inhibitor or is not administered simultaneously with an SGLT2inhibitor, the SGLT2 inhibitor and any further drug are preferablyadministered within a period of at least 2 weeks, 1 month, 2 months, 4months, 6 months or longer, e.g. 12 months or more.

In some embodiments the one or more SGLTS inhibitors, preferablycompound A, is used with co-administration with insulin, preferably asimultaneous, a sequential and/or a chronologically staggeredco-administration with insulin. Such co-administration can also be inthe form of a fixed-dose combination (FDC), e.g. a formulation includingthe one or more SGLTS inhibitors, preferably compound A, and insulincombined in a single dosage form, which is manufactured and distributedin certain respective fixed doses. In some embodiments the SGLT2inhibitor (whether used alone or in combination with another drug) isnot used in combination with1-[(3-cyano-pyridin-2-yl)methyl]-3-methyl-7-(2-butyn-1-yl)-8-[3-(R)-amino-piperidin-1-yl]-xanthineor a pharmaceutically acceptable salt thereof, i.e. the canine animal isnot treated with said compound. In some embodiments the SGLT2 inhibitoris not used in combination with a DPP-IV inhibitor, i.e., the canineanimal is not treated with a DPP-IV inhibitor.

In some embodiments, the SGLT2 inhibitor is used as a monotherapy, i.e.stand-alone therapy, i.e. no other medication is administered to thecanine animal for the treatment or prevention of the same metabolicdisorder, i.e. the metabolic disorder for which the SGLT2 inhibitor isadministered. E.g., no other medication is administered to the canineanimal for the treatment or prevention of the same metabolic disorderwithin a period of at least 2, 3, or 4 weeks before and afteradministration of the SGLT2 inhibitor.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 Urinary glucose excretion in Beagle dogs after single oral dosingof Compound A. The urine was individually sampled 0-24 h and 24-48 hafter administration. After 24 h, urine obtained by insertion of acatheter into the bladder was added to the freely sampled urine. Incontrols and the low dose (0.01 mg/kg) virtually no glucose wasdetectable in urine. Indicated p-values above bars are versus control(*, p<0.05).

FIG. 2 Sigmoidal dose response for the urinary glucose excretion in thetime period 0-24 h after administration of Compound A as shown. ED50 forglucose excretion by Compound A in Beagle dogs is 0.1 mg/kg (95% CI0.02-0.52 mg/kg).

FIG. 3 shows an X-ray powder diffraction pattern of a representativebatch of a crystalline complex of compound A with L-proline (1:1)

FIG. 4 shows a DSC/TG diagram of a representative batch of a crystallinecomplex of compound A with L-proline (1:1)

EXAMPLES

The following examples show the beneficial therapeutic effects onglycaemic control and/or insulin resistance, etc., of using one or moreSGLT2 inhibitors in canine animals, according to the present invention.These examples are intended to illustrate the invention in more detailwithout any limitation of the scope of the claims.

Example 1 Pharmacokinetics (PK) of Compound a Single Oral Dosing in Dogs

Compound A was administered to overnight fasted dogs. The groups (n=4per group) received a single administration of either oral vehicle (DIwater) containing the SGLT2 inhibitor Compound A at a dose of 1 mg/kgand 10 mg/kg or intravenous vehicle (saline) containing the SGLT2inhibitor Compound A at a dose of 1 mg/kg. PK measurements were takenuntil day 3 after a single administration of compound A or its vehicle.

TABLE 2 Pharmacokinetic data, single dose (i.v. 1 mg/kg, oral 1 and 10mg/kg) Parameter i.v. 1 mg/kg p.o. 1 mg/kg p.o. 10 mg/kg t_(max) [hour]mean 1.9 0.6 C_(max) [nmol/L] mean 4845 51525 CL [mL/min/kg] mean 0.63CL/F [mL/min/kg] mean 0.64 0.71 F [%] mean 101 92 AUC_(0→∞) mean 6702567675 616750 [nmol · h/l] T_(1/2) [hour] mean 13.4 13.9 14.5

Example 2 The Effect of Compound a on Urinary and Blood Glucose afterSingle Dosing in Dogs

Beagle dogs were fasted overnight and received a single oraladministration of Compound A at doses of 0 mg/kg b.w., 0.01 mg/kg b.w.,0.1 mg/kg b.w., or 1 mg/kg b.w. (n=3 per group) followed by a rinsingwith water (1 mL/kg b.w.)

Compound A was moistened with a small volume of a 1% (w/v) aqueousPolysorbat 80 (Tween80, Polyoxyethylene Sorbitan Monooleate, ICNBiomedicals) solution and then dissolved by slowly adding a large volumeof a 0.5% (w/v) aqueous hydroxyethylcellulose (Natrosol 250 HX,Boehringer Ingelheim) solution and stirring at room temperature forabout 15 minutes. The final concentration of Polysorbat 80 was 0.015%.Compound A was applied in a volume of 2 mL/kg. b.w.

The animals were kept individually in metabolic cages and received food2 h after administration. They had free access to water during theexperiment. Urine was collected in the time intervals 0-8 h, 8-24 h,24-32 h, and 32-48 h after administration. A catheter (Eickemeyer) wasinserted into the bladder to completely collect the 24 h urine. Thisurine was combined with the urine that had been excreted in the 8-24 hperiod. A volume of 5 mL of a 10% solution of sodium azide in saline hadbeen added to each urine collection tube before sampling. Volume ofurine was determined and samples were frozen for subsequentdetermination of glucose concentration.

During the experiment, blood samples were drawn from a forearm vein.Blood was collected in EDTA tubes prior to administration of vehicle orCompound A, and subsequently at 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 8 h, 24 h,32 h, and 48 h time points post-dose. Plasma was prepared followingblood collection and frozen for determination of glucose concentration.

-   -   A prominent increase of urinary glucose concentration and volume        was evident at the two higher doses already 8 h after        administration (FIG. 1).    -   Neither dose of compound A induced hypoglycemia, or altered the        blood glucose level in dogs as compared to normal reference        values.

In respect to urinary glucose excretion it is thus estimated that theED₅₀ is 0.1 mg/kg (FIG. 2).

Example 3 Treatment of Insulin Dependent Diabetes in Dogs

Treating Dogs with Insulin Dependent Diabetes

Insulin with the Compound A according to the invention or a combinationof active substances according to the invention, in addition toproducing an acute improvement in the glucose metabolic situation,prevents deterioration in the metabolic situation in the long term andreduces the insulin dose needed to treat the diabetic canine. This canbe observed if dogs are treated for a shorter or longer period, e.g. 2-4weeks or 3 months to 1 year, with the pharmaceutical compositionaccording to the invention and are compared to the metabolic situationprior to treatment or with dogs that have been treated with e.g. insulinalone. There is evidence of therapeutic success if daily mean bloodglucose and fructosamine level are reduced as compared to pre-treatmentlevel. Further evidence of therapeutic success is obtained if asignificantly smaller percentage of the dogs treated with apharmaceutical composition according to the invention, compared withdogs who have been treated with other medications, undergo transientdeterioration in the glucose metabolic position (e.g. hyper- orhypoglycaemia).

Example 4 Improvement of Insulin Resistance Diabetes in Female Dogs withDioestrus/Gestational Diabetes

Insulin resistance diabetes is a frequently found form of diabetes inintact female canine animals. Therapy with Compound A may be providedwith the objective of preventing the transition to manifest diabetes. Instudies over a shorter or longer period (e.g. 2-4 weeks or 1-2 years)the success of the treatment is examined by determining the fastingglucose values and/or the glucose values after a meal or after a loadingtest (intravenous glucose tolerance test or food tolerance test after adefined meal) during the study throughout the different phases of themenstrual cycle and/or after the end of the period of therapy for thestudy and comparing them with the values before the start of the studyand/or with those of a placebo group. In addition, the fructosaminevalue can be determined before and after therapy and compared with theinitial value and/or compared with dogs that have been treated withother medications or placebo. A significant drop in the fasting ornon-fasting glucose and/or fructosamine levels demonstrates the efficacyof the treatment of insulin resistance—diabetes and preventing manifestdiabetes in female dogs with a history of dioestrus/gestationaldiabetes.

Example 5 Treatment of Hyperglycaemia

In clinical studies in dogs with metabolic disorders running fordifferent lengths of time (e.g. 2 weeks to 12 months) the success of thetreatment is checked using the measurement of baseline blood glucoseand/or blood fructosamine.

The improvement of glycaemic control can furthermore be determinedestablishing diurnal blood glucose curves, e.g. a 9 or 24 hour bloodglucose curve starting prior to medication and repeated measurementspost dosing.

A significant fall in these values during or at the end of the study,compared with the initial value or compared with a placebo group, or agroup given a different therapy, proves the efficacy of a pharmaceuticalcomposition according to the invention in the reduction ofhyperglycaemia in dogs.

Alternatively, the effect of compound A on hyperglycaemia can be shownin dogs subject to a continuous glucose infusion (hyperglycaemic clamp).The normalization of the hyperglycaemia can be evaluated as compared tono treatment and/or to a combined treatment with insulin and/or atreatment with insulin alone.

Example 6 Prevention or Treatment of Hyperglycaemia AssociatedComplications

The treatment of hyperglycaemic or insulin dependent or insulinresistance diabetic dogs with Compound A according to the invention or acombination of active substances according to the invention prevents orreduces hyperglycaemia associated complications, e.g. cataractformation.

Evidence of the therapeutic success is compared with dogs that have beentreated with other antidiabetic medicaments or with placebo. The successof the treatment is determined e.g. by ophthalmological eye examinationof the development or the progression or the regression of cataractformation. And/or the time to development of a cataract and/orprogression of the cataract maturation may be determined and be comparedto dogs who have been treated with other antidiabetic medicaments orwith placebo.

Example 7 Treatment of Insulin Resistance

In clinical studies in insulin resistant dogs running for differentlengths of time (e.g. 4 weeks to 12 months) the success of the treatmentis checked using the measurement of baseline blood glucose, bloodfructosamine and blood insulin and/or c-peptide level and thecorresponding relation between the parameter in the individual dog.

Also the glucose and blood lipids (e.g. NEFA) and/or insulin valuesafter a meal or after a loading test (glucose tolerance test or insulintolerance test) during or after the end of the period of therapy for thestudy can be compared with the values before the start of the studyand/or with those of insulin resistant dogs who have been treated withother medications or placebo.

Example 8 Treatment of Pre-Diabetes in Dogs

The efficacy of SGLT2 inhibition in accordance with the invention in thetreatment of pre-diabetes characterised by pathological fasting glucoseand/or impaired glucose tolerance and/or insulin resistance can betested using clinical studies. In studies over a shorter or longerperiod (e.g. 2-4 weeks or 1-2 years) the success of the treatment isexamined by determining the fasting glucose values and/or the glucosevalues after a meal or after a loading test (intravenous glucosetolerance test or food tolerance test after a defined meal or insulintolerance test) after the end of the period of therapy for the study andcomparing them with the values before the start of the study and/or withthose of a placebo group. In addition, the fructosamine value can bedetermined before and after therapy and compared with the initial valueand/or the placebo value. A significant drop in the fasting ornon-fasting glucose and/or fructosamine levels demonstrates the efficacyof the treatment of pre-diabetes

Example 9 Effects on Body Weight, Body Composition, Dyslipidemia andDysadipokinemia

Treating dogs with metabolic disorders such as obesity, dyslipidaemia,dysadipokinemia, hepatic lipidosis, subclinical inflammation or systemicinflammation, in particular low grade systemic inflammation, which alsocomprises adipose tissue, and associated disorders, such as Syndrome X(metabolic syndrome), and/or insulin resistance, hyperglycaemia,hyperinsulinaemia, impaired glucose tolerance is also in pursuit of thegoal of preventing the transition or slowing the progression to e.g.clinically manifest consequences of the metabolic disorders e.g.hypertension, cardiomyopathy, renal dysfunction and/or musculoskeletaldisorders in canine animals.

The efficacy of a treatment can be investigated in a comparativeclinical study in which dogs are treated over a lengthy period (e.g.3-12 months) with either Compound A or a combination of activesubstances or with placebo or with a non-drug therapy (e.g. diet) orother medicaments. Prior, during and at the end of the therapy theparameter can be determined: body weight (scale) and body compositione.g. with dual-energy x-ray absorptiometry. In plasma or serum lipid(e.g. Triglycerides, Cholesterol, NEFA) and adipokine (e.g. adiponectin,leptin) profiles as well as inflammatory markers (e.g. c-reactiveprotein, monocyte chemoattractant protein-I) can be measured. Insulinand glucose level can be determined basal as well as e.g. after aloading test. Renal parameter can be determined in blood and urinarysamples (e.g. urea, creatinine, urinary albumin). Additionally, theblood pressure and/or also evidences of cardiomyopathy can beinvestigated with echocardiographic doppler ultrasound measurements. Animprovement in musculoskeletal disorders (e.g. osteoarthritis) can bequantified e.g. with activity, lameness, and pain scores.

Example 10 Preparation of1-cyano-2-(4-cyclopropyl-benzyl)-4-(β-D-glucopyranos-1-yl)-benzene(Compound A)

The following example of synthesis serves to illustrate a method ofpreparing1-cyano-2-(4-cyclopropyl-benzyl)-4-(β-D-glucopyranos-1-yl)-benzene(compound A). A method of preparing its crystalline complex withL-proline is also described. It is to be regarded only as a possiblemethod described by way of example, without restriction of the scope ofthe invention. The terms “room temperature” and “ambient temperature”are used interchangeably and denote temperatures of about 20° C. Thefollowing abbreviations are used:

-   DMF dimethylformamide-   NMP N-methyl-2-pyrrolidone-   THF tetrahydrofuran

Preparation of 4-bromo-3-hydroxymethyl-1-iodo-benzene

Oxalyl chloride (13.0 mL) is added to an ice-cold solution of2-bromo-5-iodo-benzoic acid (49.5 g) in CH₂Cl₂ (200 mL). DMF (0.2 mL) isadded and the solution is stirred at room temperature for 6 h. Then, thesolution is concentrated under reduced pressure and the residue isdissolved in THF (100 mL). The resulting solution is cooled in anice-bath and LiBH₄ (3.4 g) is added in portions. The cooling bath isremoved and the mixture is stirred at room temperature for 1 h. Thereaction mixture is diluted with THF and treated with 0.1 M hydrochloricacid. Then, the organic layer is separated and the aqueous layer isextracted with ethyl acetate. The combined organic layers are dried(Na₂SO₄) and the solvent is evaporated under reduced pressure to givethe crude product.

Yield: 47.0 g (99% of theory)

Preparation of 4-bromo-3-chloromethyl-1-iodo-benzene

Thionyl chloride (13 mL) is added to a suspension of4-bromo-3-hydroxymethyl-1-iodo-benzene (47.0 g) in dichloromethane (100mL) containing DMF (0.1 mL). The mixture is stirred at ambienttemperature for 3 h. Then, the solvent and the excess reagent is removedunder reduced pressure. The residue is triturated with methanol anddried.

Yield: 41.0 g (82% of theory)

Preparation of 4-bromo-1-iodo-3-phenoxymethyl-benzene

Phenol (13 g) dissolved in 4 M KOH solution (60 mL) is added to4-bromo-3-chloromethyl-1-iodo-benzene (41.0 g) dissolved in acetone (50mL). NaI (0.5 g) is added and the resulting mixture is stirred at 50° C.overnight. Then, water is added and the resulting mixture is extractedwith ethyl acetate. The combined extracts are dried (Na₂SO₄) and thesolvent is evaporated under reduced pressure. The residue is purified bychromatography on silica gel (cyclohexane/ethyl acetate 19:1).

Yield: 38.0 g (79% of theory)

Preparation of1-bromo-4-(1-methoxy-D-glucopyranos-1-yl)-2-(phenoxymethyl)-benzene

A 2 M solution of iPrMgCl in THF (11 mL) is added to dry LiCl (0.47 g)suspended in THF (11 mL). The mixture is stirred at room temperatureuntil all the LiCl is dissolved. This solution is added dropwise to asolution of 4-bromo-1-iodo-3-phenoxymethyl-benzene (8.0 g) intetrahydrofuran (40 mL) cooled to −60° C. under argon atmosphere. Thesolution is warmed to −40° C. and then2,3,4,6-tetrakis-O-(trimethylsilyl)-D-glucopyranone (10.7 g, 90% pure)in tetrahydrofuran (5 mL) is added. The resulting solution is warmed to−5° C. in the cooling bath and stirred for another 30 min at thistemperature. Aqueous NH₄Cl solution is added and the resultant mixtureis extracted with ethyl acetate. The combined organic extracts are driedover sodium sulfate and the solvent is removed under reduced pressure.The residue is dissolved in methanol (80 mL) and treated withmethanesulfonic acid (0.6 mL) to produce the more stable anomer solely.After stirring the reaction solution at 35-40° C. overnight, thesolution is neutralized with solid NaHCO₃ and the methanol is removedunder reduced pressure. The remainder is diluted with aqueous NaHCO₃solution and the resulting mixture is extracted with ethyl acetate. Thecombined extracts are dried over sodium sulfate and the solvent isevaporated to yield the crude product that is submitted to reductionwithout further purification.

Yield: 7.8 g (93% of theory)

Preparation of1-bromo-4-(2,3,4,6-tetra-O-acetyl-D-glucopyranos-1-yl)-2-(phenoxymethyl)-benzene

Boron trifluoride diethyletherate (4.9 mL) is added to a solution of1-bromo-4-(1-methoxy-D-glucopyranos-1-yl)-2-(phenoxymethyl)-benzene (8.7g) and triethylsilane (9.1 mL) in dichloromethane (35 mL) andacetonitrile (50 mL) cooled to −20° C. at such a rate that thetemperature maintains below −10° C. The resultant solution is warmed to0° C. over a period of 1.5 h and then treated with aqueous sodiumhydrogen carbonate solution. The resulting mixture is stirred for 0.5 h,the organic solvent is removed and the residue is extracted with ethylacetate. The combined organic layers are dried over sodium sulfate andthe solvent is removed. The residue is taken up in dichloromethane (50mL) and pyridine (9.4 mL), acetic anhydride (9.3 mL) and4-dimethylaminopyridine (0.5 g) are added in succession to the solution.The solution is stirred for 1.5 h at ambient temperature and thendiluted with dichloromethane. This solution is washed twice with 1 Mhydrochloric acid and dried over sodium sulfate. After the solvent isremoved, the residue is recrystallized from ethanol to furnish theproduct as a colourless solid.

Yield: 6.78 g (60% of theory)

Mass spectrum (ESI⁺): m/z=610/612 (Br) [M+NH₄]⁺

Preparation of2-(phenoxymethyl)-4-(2,3,4,6-tetra-O-acetyl-D-glucopyranos-1-yl)-benzonitrile

A flask charged with zinc cyanide (1.0 g), zinc (30 mg),Pd₂(dibenzylideneacetone)₃*CHCl₃ (141 mg) and tri-tert-butylphosphoniumtetrafluoroborate (111 mg) is flushed with argon. Then a solution of1-bromo-4-(2,3,4,6-tetra-O-acetyl-D-glucopyranos-1-yl)-2-(phenoxymethyl)-benzene(5.4 g) in NMP (12 mL) is added and the resulting mixture is stirred atroom temperature for 18 h. After dilution with ethyl acetate, themixture is filtered and the filtrate is washed with aqueous sodiumhydrogen carbonate solution. The organic phase is dried (sodium sulfate)and the solvent is removed. The residue is recrystallized from ethanol.

Yield: 4.10 g (84% of theory)

Mass spectrum (ESI⁺): m/z=557 [M+NH₄]⁺

Alternatively, the compound described above is synthesized starting from1-bromo-4-(2,3,4,6-tetra-O-acetyl-D-glucopyranos-1-yl)-2-(phenoxymethyl)-benzeneusing copper(I) cyanide (2 equivalents) in NMP at 210° C.

Preparation of2-bromomethyl-4-(2,3,4,6-tetra-O-acetyl-D-glucopyranos-1-yl)-benzonitrile

A 33% solution of hydrobromic acid in acetic acid (15 mL) is added to asolution of2-phenyloxymethyl-4-(2,3,4,6-tetra-O-acetyl-D-glucopyranos-1-yl)-benzonitrile(0.71 g) and acetic anhydride (0.12 mL) in acetic acid (10 ml). Theresulting solution is stirred at 55° C. for 6 h and then cooled in anice-bath. The reaction mixture is neutralized with chilled aqueouspotassium carbonate solution, and the resultant mixture is extractedwith ethyl acetate. The combined organic extracts are dried over sodiumsulfate and the solvent is removed under reduced pressure. The residueis taken up in ethyl acetate/cyclohexane (1:5), and the precipitate isseparated by filtration and dried at 50° C. to give the pure product.

Yield: 0.52 g (75% of theory)

Mass spectrum (ESI⁺): m/z=543/545 (Br) [M+NH₄]⁺

Preparation of 4-cyclopropyl-phenylboronic acid

2.5 M solution of nButyllithium in hexane (14.5 mL) is added dropwise to1-bromo-4-cyclopropyl-benzene (5.92 g) dissolved in THF (14 mL) andtoluene (50 mL) and chilled to −70° C. The resultant solution is stirredat −70° C. for 30 min before triisopropyl borate (8.5 mL) is added. Thesolution is warmed to −20° C. and then treated with 4 M aqueoushydrochloric acid (15.5 mL). The reaction mixture is further warmed toroom temperature and then the organic phase is separated. The aqueousphase is extracted with ethyl acetate and the combined organic phasesare dried (sodium sulfate). The solvent is evaporated and the residue iswashed with a mixture of ether and cyclohexane to give the product as acolourless solid.

Yield: 2.92 g (60% of theory)

Mass spectrum (ESI⁻): m/z=207 (Cl) [M+HCOO]⁻

Preparation of1-cyano-2-(4-cyclopropyl-benzyl)-4-(β-D-glucopyranos-1-yl)-benzene

An Ar filled flask is charged with2-bromomethyl-4-(2,3,4,6-tetra-O-acetyl-D-glucopyranos-1-yl)-benzonitrile(1.60 g), 4-cyclopropyl-phenylboronic acid (1.0 g), potassium carbonate(1.85 g) and a degassed 3:1 mixture of acetone and water (22 mL). Themixture is stirred at room temperature for 5 min, before it is cooled inan ice-bath. Then palladium dichloride (30 mg) is added and the reactionmixture is stirred for 16 h at ambient temperature. The mixture is thendiluted with brine and extracted with ethyl acetate. The combinedextracts are dried over sodium sulfate and the solvent is removed underreduced pressure. The residue is dissolved in methanol (20 mL) andtreated with 4 M aqueous potassium hydroxide solution (4 mL). Theresulting solution is stirred at ambient temperature for 1 h and thenneutralized with 1 M hydrochloric acid. The methanol is evaporated, andthe residue is diluted with brine and extracted with ethyl acetate. Theorganic extracts collected are dried over sodium sulfate, and thesolvent is removed. The residue is chromatographed on silica gel(dichloromethane/methanol 1:0→8:1).

Yield: 0.91 g (76% of theory)

Mass spectrum (ESI⁺): m/z=413 [M+NH₄]⁺

Preparation of a Crystalline Complex (1:1) of Compound A with L-Proline

L-proline (0.34 g) dissolved in 2.1 mL of a mixture of ethanol and water(volume ratio 10:1) is added to a solution of1-cyano-2-(4-cyclopropyl-benzyl)-4-(β-D-glucopyranos-1-yl)-benzene (1.17g, obtained as described above) dissolved in 2 mL ethanol. The resultingsolution is allowed to stand at ambient temperature. After about 16 hthe crystalline complex is isolated as white crystals by filtration. Ifnecessary the crystallisation may be initiated by scratching with aglass rod or metal spatula for example or by inoculating with seedcrystals. Residual solvent is removed by storing the crystals atslightly elevated temperature (30 to 50° C.) under vacuum for about 4 hto yield 1.27 g of the crystalline 1:1 complex of L-proline and1-cyano-2-(4-cyclopropyl-benzyl)-4-(β-D-glucopyranos-1-yl)-benzene.

Several batches of the crystalline complex according to the abovepreparation are obtained. The X-ray powder diffraction patternscoincide. The melting points are determined via DSC and evaluated asonset-temperature. Examples of melting points are approximately 89° C.,90° C., 92° C., 101° C. and 110° C. The X-ray powder diffraction patternas contained in Table 1 and as depicted in FIG. 11 and the DSC and TGdiagram in FIG. 12 correspond to a batch with a melting point ofapproximately 90° C.

The X-ray powder diffraction pattern of the crystalline complex of thecompound A and L-proline (peaks up to 30° in 2Θ) is provided above inTable 1.

Example 11 Formulations

Some examples of formulations are described in which the term “activesubstance” denotes an SGLT2 inhibitor or pharmaceutically acceptableform thereof, e.g. a prodrug or a crystalline form, for use according tothe invention. In the case of a combination with one or additionalactive substances, the term “active substance” may also include theadditional active substance.

Tablets Containing 100 mg of Active Substance

Composition:

1 tablet contains: active substance 100.0 mg lactose  80.0 mg cornstarch  34.0 mg polyvinylpyrrolidone  4.0 mg magnesium stearate  2.0 mg220.0 mgMethod of Preparation:

The active substance, lactose and starch are mixed together anduniformly moistened with an aqueous solution of thepolyvinylpyrrolidone. After the moist composition has been screened (2.0mm mesh size) and dried in a rack-type drier at 50° C. it is screenedagain (1.5 mm mesh size) and the lubricant is added. The finishedmixture is compressed to form tablets.

Weight of tablet: 220 mg

Diameter: 10 mm, biplanar, facetted on both sides and notched on oneside.

Tablets Containing 150 mg of Active Substance

Composition:

1 tablet contains: active substance 150.0 mg  powdered lactose 89.0 mgcorn starch 40.0 mg colloidal silica 10.0 mg polyvinylpyrrolidone 10.0mg magnesium stearate  1.0 mg 300.0 mg Preparation:

The active substance mixed with lactose, corn starch and silica ismoistened with a 20% aqueous polyvinylpyrrolidone solution and passedthrough a screen with a mesh size of 1.5 mm. The granules, dried at 45°C., are passed through the same screen again and mixed with thespecified amount of magnesium stearate. Tablets are pressed from themixture.

-   -   Weight of tablet: 300 mg    -   die: 10 mm, flat        Hard Gelatine Capsules Containing 150 mg of Active Substance        Composition:

1 capsule contains: active substance 150.0 mg corn starch (dried)approx. 180.0 mg lactose (powdered) approx. 87.0 mg magnesium stearate3.0 mg approx. 420.0 mgPreparation:

The active substance is mixed with the excipients, passed through ascreen with a mesh size of 0.75 mm and homogeneously mixed using asuitable apparatus. The finished mixture is packed into size 1 hardgelatine capsules.

-   -   Capsule filling: approx. 320 mg    -   Capsule shell: size 1 hard gelatine capsule.        Suppositories Containing 150 mg of Active Substance        Composition:

1 suppository contains: active substance 150.0 mg polyethyleneglycol1500 550.0 mg polyethyleneglycol 6000 460.0 mg polyoxyethylene sorbitan840.0 mg monostearate 2,000.0 mg  Preparation:

After the suppository mass has been melted the active substance ishomogeneously distributed therein and the melt is poured into chilledmoulds.

Ampoules Containing 10 mg Active Substance

Composition:

active substance 10.0 mg 0.01N hydrochloric acid/NaCl q.s.double-distilled water ad 2.0 mlPreparation:

The active substance is dissolved in the necessary amount of 0.01 N HCl,made isotonic with common salt, filtered sterile and transferred into 2ml ampoules.

Ampoules Containing 50 mg of Active Substance

Composition:

active substance 50.0 mg 0.01N hydrochloric acid/NaCl q.s.double-distilled water ad 10.0 mlPreparation:

The active substance is dissolved in the necessary amount of 0.01 N HCl,made isotonic with common salt, filtered sterile and transferred into 10ml ampoules.

REFERENCES

-   1) Beam et al. Vet. Ophtalmol. 1999. 2, 169-172-   2) Catchpole et al., Diabetologia 2005. 48: 1948-1956-   3) EP 1 213 296-   4) EP 1 354 888-   5) EP 1 344 780-   6) EP 1 489 089-   7) Nelson et al. J small Anim Pract 2000, 41, 486-490-   8) Verkest, Vet J, in press doi.org/10.1016/j.tvjl.2013.09.057-   9) Wang et al. J Diabet. Compl. in press,    doi:0.1016/j.jdiacomp.2013.11.002-   10) WO 01/27128-   11) WO 03/099836-   12) WO 2004/007517-   13) WO 2004/080990-   14) WO 2005/012326-   15) WO 2005/092877-   16) WO 2006/034489-   17) WO 2006/064033-   18) WO 2006/117359-   19) WO 2006/117360-   20) WO 2006/120208-   21) WO 2007/025943-   22) WO 2007/028814-   23) WO 2007/031548-   24) WO 2007/093610-   25) WO 2007/114475-   26) WO 2007/128749-   27) WO 2007/140191-   28) WO 2008/002824-   29) WO 2008/013280-   30) WO 2008/042688-   31) WO 2008/049923-   32) WO 2008/055870-   33) WO 2008/055940-   34) WO 2008/069327-   35) WO 2008/116179-   36) WO 2009/014970-   37) WO 2009/022008-   38) WO 2009/022020-   39) WO 2009/035969-   40) WO 2010/023594-   41) WO 2011/039107-   42) WO 2011/039108-   43) WO 2011/117295-   44) WO 2014/016381

The invention claimed is:
 1. A method for treating a metabolic disorderin a canine animal, the method comprising administering to the canineanimal one or more active ingredients, the one or more activeingredients consisting of: a single SGLT2 inhibitor or pharmaceuticalacceptable form thereof, wherein the single SGLT2 inhibitor is a1-cyano-2-(4-cyclopropyl-benzyl)-4-(β-D-glucopyranos-1-yl)-benzenerepresented by the following formula:

or a combination of the SGLT2 inhibitor or pharmaceutical acceptableform thereof and insulin; wherein: the metabolic disorder is selectedfrom the group consisting of: ketoacidosis, pre-diabetes, insulindependent diabetes mellitus, insulin resistance diabetes, insulinresistance, obesity, hyperglycemia, hyperglycemia induced cataractformation, impaired glucose tolerance, hyperinsulinemia, dyslipidemia,dysadipokinemia, subclinical inflammation, systemic inflammation, lowgrade systemic inflammation, hepatic lipidosis, inflammation of thepancreas, metabolic disorder consequences, Syndrome X (metabolicsyndrome), and combinations thereof; and the SGLT2 inhibitor orpharmaceutically acceptable form thereof is administered at a dose of0.01 to 1.0 mg/kg body weight per day.
 2. A method for treating ametabolic disorder in a canine animal, the method comprisingadministering to the canine animal one or more active ingredients, theone or more active ingredients consisting of: a single SGLT2 inhibitoror pharmaceutical acceptable form thereof, wherein the single SGLT2inhibitor is a1-cyano-2-(4-cyclopropyl-benzyl)-4-(β-D-glucopyranos-1-yl)-benzenerepresented by the following formula:

or a combination of the SGLT2 inhibitor or pharmaceutical acceptableform thereof and insulin; wherein: the metabolic disorder is selectedfrom the group consisting of: ketoacidosis, pre-diabetes, insulindependent diabetes mellitus, insulin resistance diabetes, insulinresistance, obesity, hyperglycemia, hyperglycemia induced cataractformation, impaired glucose tolerance, hyperinsulinemia, dyslipidemia,dysadipokinemia, subclinical inflammation, systemic inflammation, lowgrade systemic inflammation, hepatic lipidosis, inflammation of thepancreas, metabolic disorder consequences, Syndrome X (metabolicsyndrome), and combinations thereof; wherein the treatment includesprevention or remission of hyperglycemia induced cataract formation. 3.The method of claim 1, wherein the treatment includes slowed progressionor remission of the metabolic disorder consequences, and the metabolicdisorder consequences are selected from the group consisting ofhypertension, renal dysfunction or musculoskeletal disorders andcombinations thereof.
 4. The method of claim 1, wherein the metabolicdisorder is selected from clinical conditions associated withpre-diabetes, insulin dependent diabetes mellitus, insulin resistance,or combinations thereof.
 5. The method of claim 4, wherein said clinicalconditions are selected from the group consisting of ketoacidosis,insulin resistance, obesity, hyperglycemia, hyperglycemia inducedcataract formation, impaired glucose tolerance, hyperinsulinemia,dyslipidemia, dysadipokinemia, subclinical inflammation, systemicinflammation, low grade systemic inflammation, hepatic lipidosis,inflammation of the pancreas, metabolic disorder consequences, such ashypertension, renal dysfunction and/or musculoskeletal disorders,Syndrome X (metabolic syndrome) and combinations thereof.
 6. The methodof claim 1, wherein said metabolic disorder is a metabolic disorderconsequence selected from the group consisting of hypertension, renaldysfunction a musculoskeletal disorder, and combinations thereof.
 7. Themethod of claim 1, wherein the canine animal is suffering from diabetes,pre-diabetes or insulin dependent diabetes.
 8. The method of claim 7,wherein the canine animal is a dog.
 9. The method of claim 1, whereinthe pharmaceutically acceptable form thereof is a crystalline complexbetween the SGLT2 inhibitor or pharmaceutically acceptable form thereofand an amino acid.
 10. The method of claim 9, wherein said amino acid isproline.
 11. The method of claim 10, wherein said amino acid isL-proline.
 12. The method of claim 1, wherein the SGLT2 inhibitor orpharmaceutically acceptable form thereof is administered orally orparenterally.
 13. The method of claim 1, wherein the SGLT2 inhibitor orpharmaceutically acceptable form thereof is administered orally.
 14. Themethod of claim 1, wherein the SGLT2 inhibitor or pharmaceuticallyacceptable form thereof is administered once per day.
 15. The method ofclaim 1, wherein the SGLT2 inhibitor or pharmaceutically acceptable formthereof is administered in combination with insulin.
 16. The method ofclaim 15, wherein administration of the combination is a simultaneous, asequential, or a chronologically staggered co-administration.
 17. Themethod of claim 15, wherein administration of the combination is achronologically staggered combination with a long acting insulin. 18.The method according to claim 1, wherein the composition comprises a 1:1crystalline complex of the SGLT2 inhibitor or pharmaceuticallyacceptable form thereof and an amino acid, and the crystalline complexis a crystalline hydrate.
 19. The method according to claim 1, whereinthe composition comprises a 1:1:1 crystalline complex of the SGLT2inhibitor or pharmaceutically acceptable form thereof, L-proline andwater in a crystalline form.
 20. The method of claim 1, wherein theSGLT2 inhibitor or pharmaceutically acceptable form thereof isadministered at a dose of 0.03 to 0.3 mg/kg body weight per day.
 21. Themethod according to claim 1, wherein the SGLT2 inhibitor orpharmaceutically acceptable form thereof is administered at a dose of0.01 to 0.5 mg/kg body weight per day.
 22. A method of treatment of ametabolic disorder in a canine animal comprising administering to thecanine animal a composition comprising a1-cyano-2-(4-cyclopropyl-benzyl)-4-(β-D-glucopyranos-1-yl)-benzenerepresented by the following formula:

wherein: the metabolic disorder is selected from the group consistingof: ketoacidosis, pre-diabetes, insulin dependent diabetes mellitus,insulin resistance diabetes, insulin resistance, obesity, hyperglycemia,hyperglycemia induced cataract formation, impaired glucose tolerance,hyperinsulinemia, dyslipidemia, dysadipokinemia, subclinicalinflammation, systemic inflammation, low grade systemic inflammation,hepatic lipidosis, inflammation of the pancreas, metabolic disorderconsequences, Syndrome X (metabolic syndrome), and combinations thereof;and the composition comprises a 1:1 crystalline complex of the SGLT2inhibitor or pharmaceutically acceptable form thereof and an amino acid,and the crystalline complex is a crystalline hydrate.